Bax agonist, compositions, and methods related thereto

ABSTRACT

The disclosure relates to BAX activators and therapeutic uses relates thereto. In certain embodiments, the disclosure relates to methods of treating or preventing cancer, such as lung cancer, comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a compound disclosed herein or pharmaceutically acceptable salt to a subject in need thereof.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No.61/525,249 filed on 19 Aug. 2011 and U.S. Provisional Application No.61/648,887 filed 18 May 2012, both hereby incorporated by reference intheir entirety.

ACKNOWLEDGEMENT

This invention was made with government support under Grant R01CA160489awarded by the National Institutes of Health. The government has certainrights in the invention.

BACKGROUND

BAX, a member of the BCL-2 (B-cell lymphoma-2) family, is anuclear-encoded protein that is able to pierce the mitochondrial outermembrane to mediate cell death by apoptosis. BAX adopts a globularα-helical structure and converts into pore-forming protein by changingconformation and assembling into oligomeric complexes in themitochondrial outer membrane. Proteins from the mitochondrialintermembrane space then empty into the cytosol to activate proteasesthat degrade the cell.

Cancer cells are able to evade apoptosis by the dysregulation of pro-and anti-apoptotic Bcl-2 family proteins. The expression of BAX appearsto play an important role in suppressing cancer development anddecreased BAX levels contribute to chemoresistance in a number ofcancers, including, but not limited to, lung cancer, chronic lymphocyticleukemia (CLL), and prostate cancer, and. See Xin & Deng, J Biol Chem.,(2005), 280, 10781-10789; Pepper et al., Br J Cancer, (1997) 76: 935-8.Because BAX is extensively expressed in both small cell lung cancer andnon-small cell lung cancer cells, BAX agonists could be particularlyuseful for treating lung cancer. Thus, there is a need to identifycompounds that activate BAX.

SUMMARY

This disclosure relates to BAX activators and therapeutic uses relatesthereto. In certain embodiments, the disclosure relates to methods oftreating or preventing cancer, such as lung cancer, comprisingadministering a therapeutically effective amount of a pharmaceuticalcomposition comprising a compound disclosed herein or pharmaceuticallyacceptable salt to a subject in need thereof. In certain embodiments,the disclosure relates to compounds or derivatives, prodrugs, or estersof compounds disclosed herein optionally substituted with one or moresubstituents.

In certain embodiments, the disclosure relates to compounds of FormulaI,

or salt thereof wherein,

is a double or single bond;

A ring is a carbocyclyl, aryl, or heterocyclyl;

X is CH or N;

Y is (CH₂)_(n) or a direct bond to the A ring, wherein n is 1 or 2;

R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are each individually and independentlyhydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino,(alkyl)₂amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,aryl, or heterocyclyl, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areoptionally substituted with one or more, the same or different, R¹⁰;

R² is nitro or amino wherein R² is optionally substituted with one ormore, the same or different, R¹⁰;

R⁹ is hydroxy, alkoxy, or amino, wherein R⁹ is optionally substitutedwith one or more, the same or different, R¹⁰;

R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹⁰ is optionally substituted with one or more,the same or different, R¹¹;

R¹¹ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹¹ is optionally substituted with one or more,the same or different, R¹²;

R¹² is halogen, nitro, cyano, hydroxy, trifluoromethoxy,trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl,methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino,ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino,acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio,methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,carbocyclyl, aryl, or heterocyclyl.

In certain embodiments, R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are hydrogen and

is a double bond.

In certain embodiments, A ring is phenyl ortho-, meta- orpara-substituted with R⁹ wherein R⁹ is hydroxy, alkoxy, alkylamino, orsubstituted with hydroxy, (alkyl)₂amino, alkylsulfamoyl,dialkylsulfamoyl, or a heterocyclyl such as pyrrolidinyl, morpholinyl,piperazinyl, wherein heterocyclyl may be substituted with one or moreR¹², Y is a direct bond to the A ring, X is CH, R² is nitro, amino,amide, urea, or sulfonamide wherein R² is substituted with one or moreR¹¹.

In certain embodiments, the A ring is aryl or heterocyclyl such aspyridinyl ortho- or meta- or para-substituted with R⁹ and R² is nitro.

In certain embodiments, R² is amide, urea, or sulfonamide substitutedwith alkyl or aryl, and R⁹ is hydroxyl.

In certain embodiments,

is a double bond, X is N, and R⁹ is alkoxy.

In certain embodiments,

is a single bond, Y is a direct bond to the A ring.

In certain embodiments,

is a double bond, Y is (CH₂)_(n) wherein n is 1.

In certain embodiments, the disclosure relates to pharmaceuticalcompositions comprising compounds disclosed herein such as those ofFormula I, IA, IB, or II or pharmaceutically acceptable salts and apharmaceutically acceptable excipient. In certain embodiments thepharmaceutical compositions further comprising a second therapeuticagent.

In certain embodiments, the disclosure relates to methods of treating orpreventing cancer comprising administering a pharmaceutical compositioncomprising compounds disclosed herein such as those of Formula I, IA,IB, or II to a subject diagnosed with, exhibiting symptoms of, or atrisk of cancer. In certain embodiments, the cancer is selected from thegroup consisting of leukemia, melanoma, cervical, ovarian, colon,breast, gastric, lung, skin, ovarian, pancreatic, prostate, head, neck,and renal cancer. In certain embodiments, the pharmaceutical compositionis administered in combination with a second chemotherapeutic agent suchas, but not limited to, gefitinib, erlotinib, docetaxel, cis-platin,5-fluorouracil, gemcitabine, tegafur, raltitrexed, methotrexate,cytosine arabinoside, hydroxyurea, adriamycin, bleomycin, doxorubicin,daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin andmithramycin, vincristine, vinblastine, vindesine, vinorelbine,paclitaxel, taxotere, etoposide, teniposide, amsacrine, topotecan,camptothecin bortezomib anegrilide, tamoxifen, toremifene, raloxifene,droloxifene, iodoxyfene fulvestrant, bicalutamide, flutamide,nilutamide, cyproterone, goserelin, leuprorelin, buserelin, megestrolanastrozole, letrozole, vorazole, exemestane, finasteride, marimastat,trastuzumab, cetuximab, dasatinib, imatinib, bevacizumab,combretastatin, thalidomide, and/or lenalidomide or combinationsthereof.

In certain embodiments, the disclosure relates to therapeutic methodsdisclosed herein wherein the pharmaceutical compositions areadministered before, after or during radiotherapy.

In certain embodiments, the disclosure relates to uses of compoundsdisclosed herein in the production of a medicament for the treatment orprevention of cancer.

In certain embodiments, the disclosure relates to methods of preparingcompounds disclosed herein comprising mixing starting materials andreagents disclosed herein under conditions that the compounds areformed.

In certain embodiments, the disclosure relates to methods of inhibitingphosphorylation of BAX at Ser184.

In some embodiments, the disclosure relates to methods of testingcompounds for the ability to inhibit BAX phosphorylation comprisingmixing a compound and a BAX protein and assaying for phosphorylation atSer184 by comparing the ability of nicotine to phosphorylate Ser184after exposing BAX to a test compound.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows data suggesting phosphorylation of BAX with nicotine at Ser184 inactivates the proapoptotic function of BAX. (A) A549 cellsexpressing endogenous BAX were metabolically labeled with³²P-orthophosphoric acid and treated with nicotine for 60 min. BAX wasimmunoprecipitated by using an agarose-conjugated BAX antibody.Phosphorylation of BAX was determined by autoradiography. (B)Phosphoamino acid analysis was performed using the phosphorylated BAXinduced by nicotine. (C) A549 cells were treated with cisplatin (40 μM)in the absence or presence of nicotine (1 μM) for various times asindicated. Cell viability was analyzed for Annexin-V and PI binding byflow cytometry. (D) The pcDNA3 plasmids bearing GFP-WT, GFP-S184A orGFP-184E were transfected into H157 cells. Cells were metabolicallylabeled with ³²P-orthophosphoric acid and treated with nicotine for 60min. Phosphorylated GFP-tagged BAX was analyzed by autoradiography. (E)The pcDNA3 plasmids bearing GFP-WT, GFP-S184A or GFP-184E weretransfected into H157 cells. After 48 h, cells were treated withcisplatin (Cis) in the absence or presence of nicotine for 24 h. Cellviability was analyzed as in (C).

FIG. 2 shows data on the effect of small molecules that structurallytarget the Ser184 site of BAX on apoptosis of human lung cancer cells.(A) Expression of BAX or Bcl2 in various lung cancer cell lines orprimary normal small airway epithelial cells (SAEC) was analyzed byWestern blot. (B) H1299, A549 or SAEC cells were treated with varioustypes of small molecules (1 μM) for 48 h. Cell viability was assessedusing ApoAlert Annexin-V kit. DMSO or cisplatin (40 μM) was used as anegative or positive control, respectively.

FIG. 3 shows data suggesting in vivo anti-tumor activity of2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol. (A) A549 lung cancerxenografts were administered intraperitoneally (q.d.) with vehiclecontrol or 2-(2-Nitro-fluoren-9-ylidenemethyl)phenol (SMBA1) asindicated doses. At day 6, tumors were removed and photographed. (B)A549 lung cancer xenograft mice were treated with vehicle control orincreasing doses of 2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol (i.e. 25mg/kg, 50 mg/kg, or 75 mg/kg) for 14 days. Tumor volume was estimated bycaliper measurements, data are mean±s.e.m. (C) Mice with xenografts weretreated with vehicle or 2-(2-Nitro-fluoren-9-ylidenemethyl)phenol (50mg/kg) for 24 h. Apoptosis in tumor tissues was measured by TUNEL assay.

FIG. 4 schematically illustrates methods for preparing compoundsdisclosed herein.

FIGS. 5A-C show data from the sulforhodamine B (SRB) assay forsuppressing lung cancer growth.

FIG. 6 shows in vivo anti-lung cancer activity of SMBA1 and its analog(CYD-2-11).

FIG. 7 shows activity data for embodiments in A549 lung cancer parentalcells (A549-P) and radiation resistant cells (A549-IRR).

DETAILED DESCRIPTION

BAX is a Bcl-2 family protein. Human BAX isoform alpha has an amino acidsequence of MDGSGEQPRG GGPTSSEQIM KTGALLLQGF IQDRAGRMGG EAPELALDPVPQDASTKKLS ECLKRIGDEL DSNMELQRMI AAVDTDSPRE VFFRVAADMF SDGNFNWGRVVALFYFASKL VLKALCTKVP ELIRTIMGWT LDFLRERLLG WIQDQGGWDG LLSYFGTPTWQTVTIFVAGV LTASLTIWKK MG (SEQ ID NO:1). A pocket is located in thehydrophobic C-terminal tail of BAX, which regulates the subcellularlocation and its ability to insert into mitochondrial membranes.Phosphorylation or dephosphorylation of BAX at Ser184 negatively orpositively regulates the proapoptotic activity of BAX. Ser184 residuewas chosen as a docking site for screening of small molecules thatactivate BAX using the computerized DOCK suite of programs and adatabase of 300,000 small molecules from the National Cancer Institute(NCI) filtered to follow the Lipinski rules. It has been discovered thatcertain compounds activate BAX. Thus, in certain embodiments, thedisclosure relates to compounds disclosed herein, salts, substitutedforms, and derivatives. In certain embodiments, the disclosurecontemplates pharmaceutical compositions containing these compounds foruse in the treatment or prevent of BAX related diseases or conditionssuch as cancer.

Compounds

In certain embodiments, the disclosure contemplates compounds asprovided for in Formula I below,

or salts thereof wherein,

is a double or single bond;

A ring is a carbocyclyl, aryl, or heterocyclyl;

X is CH or N;

Y is (CH₂)_(n) or a direct bond to the A ring, wherein n is 1 or 2;

R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are each individually and independentlyhydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino,(alkyl)₂amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,aryl, or heterocyclyl, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areoptionally substituted with one or more, the same or different, R¹⁰;

R² is nitro or amino wherein R² is optionally substituted with one ormore, the same or different, R¹⁰;

R⁹ is hydroxy, alkoxy, or amino, wherein R⁹ is optionally substitutedwith one or more, the same or different, R¹⁰;

R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹⁰ is optionally substituted with one or more,the same or different, R¹¹;

R¹¹ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹¹ is optionally substituted with one or more,the same or different, R¹²;

R¹² is halogen, nitro, cyano, hydroxy, trifluoromethoxy,trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl,methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino,ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino,acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio,methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,carbocyclyl, aryl, or heterocyclyl.

In certain embodiments, R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are hydrogen.

In certain embodiments,

is a double bond.

In certain embodiments, A ring is phenyl ortho-, meta- orpara-substituted with R⁹ wherein R⁹ is hydroxy, alkoxy, alkylamino, orsubstituted with hydroxy, (alkyl)₂amino, alkylsulfamoyl,dialkylsulfamoyl, or a heterocyclyl such as pyrrolidinyl, morpholinyl,piperazinyl, wherein heterocyclyl may be substituted with one or moreR¹².

In certain embodiments, Y is a direct bond to the A ring.

In certain embodiments, X is CH.

In certain embodiments, R² is nitro, amino, amide, urea, or sulfonamidewherein R² is substituted with one or more R¹¹.

In certain embodiments, the A ring is an aryl or heterocyclyl such aspyridinyl ortho- or meta- or para-substituted with R⁹.

In certain embodiments, R² is nitro.

In certain embodiments, X is N.

In certain embodiments, R⁹ is alkoxy.

In certain embodiments, Y is a direct bond to the A ring.

In certain embodiments, Y is (CH₂)_(n) wherein n is 1.

In certain embodiments, the compounds of Formula I have Formula IA,

or salts thereof wherein,

R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are each individually and independentlyhydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino,(alkyl)₂amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,aryl, or heterocyclyl, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areoptionally substituted with one or more, the same or different, R¹⁰;

R² is nitro or amino optionally substituted with one or more, the sameor different, R¹⁰;

R⁹ is hydroxy, alkoxy, or amino, wherein R⁹ is optionally substitutedwith one or more, the same or different, R¹⁰;

R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹⁰ is optionally substituted with one or more,the same or different, R¹¹;

R¹¹ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹¹ is optionally substituted with one or more,the same or different, R¹²;

R¹² is halogen, nitro, cyano, hydroxy, trifluoromethoxy,trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl,methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino,ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino,acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio,methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,carbocyclyl, aryl, or heterocyclyl.

In certain embodiments, the compounds of Formula I have Formula IB,

or salts thereof wherein,

Z is O, S, CH₂, or NH;

W is hydroxy, amino, alkylamino, dialkylamino, aryl, or heterocyclylwherein W is optionally substituted with one or more R¹¹;

R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are each individually and independentlyhydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino,(alkyl)₂amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,aryl, or heterocyclyl, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areoptionally substituted with one or more, the same or different, R¹⁰;

R² is nitro or amino optionally substituted with one or more, the sameor different, R¹⁰;

R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹⁰ is optionally substituted with one or more,the same or different, R¹¹;

R¹¹ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹¹ is optionally substituted with one or more,the same or different, R¹²;

R¹² is halogen, nitro, cyano, hydroxy, trifluoromethoxy,trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl,methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino,ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino,acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio,methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,carbocyclyl, aryl, or heterocyclyl.

In certain embodiments, the compounds of Formula I have Formula IC,

or salts thereof wherein,

R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are each individually and independentlyhydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino,(alkyl)₂amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,aryl, or heterocyclyl, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areoptionally substituted with one or more, the same or different, R¹⁰;

R² is nitro or amino optionally substituted with one or more, the sameor different, R¹⁰;

R⁹ is hydroxy, alkoxy, or amino, wherein R⁹ is optionally substitutedwith one or more, the same or different, R¹⁰;

R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹⁰ is optionally substituted with one or more,the same or different, R¹¹;

R¹¹ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹¹ is optionally substituted with one or more,the same or different, R¹²;

R¹² is halogen, nitro, cyano, hydroxy, trifluoromethoxy,trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl,methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino,ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino,acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio,methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,carbocyclyl, aryl, or heterocyclyl.

In certain embodiments, the compounds of Formula I have Formula ID,

or salts thereof wherein,

Z is O, S, CH₂, or NH;

W is hydroxy, amino, alkylamino, dialkylamino, aryl, or heterocyclylwherein W is optionally substituted with one or more R¹¹;

R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are each individually and independentlyhydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino,(alkyl)₂amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,aryl, or heterocyclyl, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areoptionally substituted with one or more, the same or different, R¹⁰;

R² is nitro or amino optionally substituted with one or more, the sameor different, R¹⁰;

R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹⁰ is optionally substituted with one or more,the same or different, R¹¹;

R¹¹ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹¹ is optionally substituted with one or more,the same or different, R¹²;

R¹² is halogen, nitro, cyano, hydroxy, trifluoromethoxy,trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl,methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino,ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino,acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio,methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,carbocyclyl, aryl, or heterocyclyl.

In certain embodiments, the disclosure relates to compounds of FormulaII,

or salts thereof wherein,

U is —C(═O)— or —SO₂—;

R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are each individually and independentlyhydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino,(alkyl)₂amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,aryl, or heterocyclyl, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areoptionally substituted with one or more, the same or different, R¹⁰;

R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹⁰ is optionally substituted with one or more,the same or different, R¹¹;

R¹¹ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹¹ is optionally substituted with one or more,the same or different, R¹²;

R¹² is halogen, nitro, cyano, hydroxy, trifluoromethoxy,trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl,methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino,ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino,acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio,methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,carbocyclyl, aryl, or heterocyclyl.

In certain embodiments, the compounds as provided for in Formula Icbelow are provided,

or salts thereof wherein,

is a double or single bond;

A ring is a carbocyclyl, aryl, or heterocyclyl;

X is CH or N;

Y is (CH₂)_(n) or a direct bond to the A ring, wherein n is 1 or 2;

R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are each individually and independentlyhydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino,(alkyl)₂amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,aryl, or heterocyclyl, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areoptionally substituted with one or more, the same or different, R¹⁰;

R² is nitro or amino wherein R² is optionally substituted with one ormore, the same or different, R¹⁰;

R⁹ is hydroxy, alkoxy, amino, halo, a heterocycle (such as piperazine),or an amide, urea or sulfonamide, wherein R⁹ is optionally substitutedwith one or more, the same or different, R¹⁰;

R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹⁰ is optionally substituted with one or more,the same or different, R¹¹;

R¹¹ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)₂amino,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, orheterocyclyl, wherein R¹¹ is optionally substituted with one or more,the same or different, R¹²;

R¹² is halogen, nitro, cyano, hydroxy, trifluoromethoxy,trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl,methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino,ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino,acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio,methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,carbocyclyl, aryl, or heterocyclyl.

In certain embodiments, R⁹ is halogen. In certain other embodiments, R⁹is a heterocycle.

Combination Therapies

The cancer treatments disclosed herein can be applied as a sole therapyor can involve, conventional surgery or radiotherapy or chemotherapy.Such chemotherapy can include one or more of the following categories ofanti-tumor agents:

(i) antiproliferative/antineoplastic drugs and combinations thereof, asused in medical oncology, such as alkylating agents (for examplecis-platin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan,chlorambucil, busulfan and nitrosoureas); antimetabolites (for exampleantifolates such as fluoropyrimidines like 5-fluorouracil andgemcitabine, tegafur, raltitrexed, methotrexate, cytosine arabinosideand hydroxyurea); antitumor antibiotics (for example anthracyclines likeadriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin,mitomycin-C, dactinomycin and mithramycin); antimitotic agents (forexample vinca alkaloids like vincristine, vinblastine, vindesine andvinorelbine and taxoids like paclitaxel and taxotere); and topoisomeraseinhibitors (for example epipodophyllotoxins like etoposide andteniposide, amsacrine, topotecan and camptothecin); and proteosomeinhibitors (for example bortezomib [Velcade®]); and the agent anegrilide[Agrylin®]; and the agent alpha-interferon

(ii) cytostatic agents such as antioestrogens (for example tamoxifen,toremifene, raloxifene, droloxifene and iodoxyfene), oestrogen receptordown regulators (for example fulvestrant), antiandrogens (for examplebicalutamide, flutamide, nilutamide and cyproterone acetate), LHRHantagonists or LHRH agonists (for example goserelin, leuprorelin andbuserelin), progestogens (for example megestrol acetate), aromataseinhibitors (for example as anastrozole, letrozole, vorazole andexemestane) and inhibitors of 5α-reductase such as finasteride;

(iii) agents which inhibit cancer cell invasion (for examplemetalloproteinase inhibitors like marimastat and inhibitors of urokinaseplasminogen activator receptor function);

(iv) inhibitors of growth factor function, for example such inhibitorsinclude growth factor antibodies, growth factor receptor antibodies (forexample the anti-Her2 antibody trastuzumab and the anti-epidermal growthfactor receptor (EGFR) antibody, cetuximab), farnesyl transferaseinhibitors, tyrosine kinase inhibitors and serine/threonine kinaseinhibitors, for example inhibitors of the epidermal growth factor familyfor example EGFR family tyrosine kinase inhibitors such as:N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine(gefitinib),N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine(erlotinib), and6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)quinazolin-4-amine(CI 1033), for example inhibitors of the platelet-derived growth factorfamily and for example inhibitors of the hepatocyte growth factorfamily, for example inhibitors of phosphotidylinositol 3-kinase (PI3K)and for example inhibitors of mitogen activated protein kinase kinase(MEK1/2) and for example inhibitors of protein kinase B (PKB/Akt), forexample inhibitors of Src tyrosine kinase family and/or Abelson (AbI)tyrosine kinase family such as dasatinib (BMS-354825) and imatinibmesylate (Gleevec™); and any agents that modify STAT signalling;

(v) antiangiogenic agents such as those which inhibit the effects ofvascular endothelial growth factor, (for example the anti-vascularendothelial cell growth factor antibody bevacizumab [Avastin™]) andcompounds that work by other mechanisms (for example linomide,inhibitors of integrin ocvβ3 function and angiostatin);

(vi) vascular damaging agents such as Combretastatin A4;

(vii) antisense therapies, for example those which are directed to thetargets listed above, such as an anti-RAS antisense; and

(viii) immunotherapy approaches, including for example ex-vivo andin-vivo approaches to increase the immunogenicity of patient tumorcells, such as transfection with cytokines such as interleukin 2,interleukin 4 or granulocyte-macrophage colony stimulating factor,approaches to decrease T-cell anergy, approaches using transfectedimmune cells such as cytokine-transfected dendritic cells, approachesusing cytokine-transfected tumor cell lines and approaches usinganti-idiotypic antibodies, and approaches using the immunomodulatorydrugs thalidomide and lenalidomide [Revlimid®].

Formulations

Pharmaceutical compositions disclosed herein can be in the form ofpharmaceutically acceptable salts, as generally described below. Somepreferred, but non-limiting examples of suitable pharmaceuticallyacceptable organic and/or inorganic acids are hydrochloric acid,hydrobromic acid, sulfuric acid, nitric acid, acetic acid and citricacid, as well as other pharmaceutically acceptable acids known per se(for which reference is made to the references referred to below).

When the compounds of the disclosure contain an acidic group as well asa basic group, the compounds of the disclosure can also form internalsalts, and such compounds are within the scope of the disclosure. When acompound contains a hydrogen-donating heteroatom (e.g. NH), salts arecontemplated to cover isomers formed by transfer of the hydrogen atom toa basic group or atom within the molecule.

Pharmaceutically acceptable salts of the compounds include the acidaddition and base salts thereof. Suitable acid addition salts are formedfrom acids which form non-toxic salts. Examples include the acetate,adipate, aspartate, benzoate, besylate, bicarbonate/carbonate,bisulphate/sulphate, borate, camsylate, citrate, cyclamate, edisylate,esylate, formate, fumarate, gluceptate, gluconate, glucuronate,hexafluorophosphate, hibenzate, hydrochloride/chloride,hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate,maleate, malonate, mesylate, methylsulphate, naphthylate, 2-napsylate,nicotinate, nitrate, orotate, oxalate, palmitate, pamoate,phosphate/hydrogen phosphate/dihydrogen phosphate, pyroglutamate,saccharate, stearate, succinate, tannate, tartrate, tosylate,trifluoroacetate and xinofoate salts. Suitable base salts are formedfrom bases which form non-toxic salts. Examples include the aluminium,arginine, benzathine, calcium, choline, diethylamine, diolamine,glycine, lysine, magnesium, meglumine, olamine, potassium, sodium,tromethamine and zinc salts. Hemisalts of acids and bases can also beformed, for example, hemisulphate and hemicalcium salts. For a review onsuitable salts, see Handbook of Pharmaceutical Salts: Properties,Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002), incorporatedherein by reference.

The compounds described herein can be administered in the form ofprodrugs. A prodrug can include a covalently bonded carrier whichreleases the active parent drug when administered to a mammaliansubject. Prodrugs can be prepared by modifying functional groups presentin the compounds in such a way that the modifications are cleaved,either in routine manipulation or in vivo, to the parent compounds.Prodrugs include, for example, compounds wherein a hydroxyl group isbonded to any group that, when administered to a mammalian subject,cleaves to form a free hydroxyl group. Examples of prodrugs include, butare not limited to, acetate, formate and benzoate derivatives of alcoholfunctional groups in the compounds. Examples of structuring a compoundas prodrugs can be found in the book of Testa and Caner, Hydrolysis inDrug and Prodrug Metabolism, Wiley (2006) hereby incorporated byreference. Typical prodrugs form the active metabolite by transformationof the prodrug by hydrolytic enzymes, the hydrolysis of amides, lactams,peptides, carboxylic acid esters, epoxides or the cleavage of esters ofinorganic acids.

Pharmaceutical compositions typically comprise an effective amount of acompound and a suitable pharmaceutical acceptable carrier. Thepreparations can be prepared in a manner known per se, which usuallyinvolves mixing the at least one compound according to the disclosurewith the one or more pharmaceutically acceptable carriers, and, ifdesired, in combination with other pharmaceutical active compounds, whennecessary under aseptic conditions. Reference is made to U.S. Pat. No.6,372,778, U.S. Pat. No. 6,369,086, U.S. Pat. No. 6,369,087 and U.S.Pat. No. 6,372,733 and the further references mentioned above, as wellas to the standard handbooks, such as the latest edition of Remington'sPharmaceutical Sciences. It is well known that ester prodrugs arereadily degraded in the body to release the corresponding alcohol. Seee.g., Imai, Drug Metab Pharmacokinet (2006) 21(3):173-85, entitled“Human carboxylesterase isozymes: catalytic properties and rational drugdesign.”

Generally, for pharmaceutical use, the compounds can be formulated as apharmaceutical preparation comprising at least one compound and at leastone pharmaceutically acceptable carrier, diluent or excipient and/oradjuvant, and optionally one or more further pharmaceutically activecompounds.

The pharmaceutical preparations of the disclosure are preferably in aunit dosage form, and can be suitably packaged, for example in a box,blister, vial, bottle, sachet, ampoule or in any other suitablesingle-dose or multi-dose holder or container (which can be properlylabeled); optionally with one or more leaflets containing productinformation and/or instructions for use. Generally, such unit dosageswill contain between 1 and 1000 mg, and usually between 5 and 500 mg, ofthe at least one compound of the disclosure e.g., about 10, 25, 50, 100,200, 300 or 400 mg per unit dosage.

The compounds can be administered by a variety of routes including theoral, ocular, rectal, transdermal, subcutaneous, intravenous,intramuscular or intranasal routes, depending mainly on the specificpreparation used. The compound will generally be administered in an“effective amount,” by which it is meant any amount of a compound that,upon suitable administration, is sufficient to achieve the desiredtherapeutic or prophylactic effect in the subject to which it isadministered. Usually, depending on the condition to be prevented ortreated and the route of administration, such an effective amount willusually be between 0.01 to 1000 mg per kilogram body weight of thepatient per day, more often between 0.1 and 500 mg, such as between 1and 250 mg, for example about 5, 10, 20, 50, 100, 150, 200 or 250 mg,per kilogram body weight of the patient per day, which can beadministered as a single daily dose, divided over one or more dailydoses. The amount(s) to be administered, the route of administration andthe further treatment regimen can be determined by the treatingclinician, depending on factors such as the age, gender and generalcondition of the patient and the nature and severity of thedisease/symptoms to be treated. Reference is made to U.S. Pat. No.6,372,778, U.S. Pat. No. 6,369,086, U.S. Pat. No. 6,369,087 and U.S.Pat. No. 6,372,733 and the further references mentioned above, as wellas to the standard handbooks, such as the latest edition of Remington'sPharmaceutical Sciences.

Formulations containing one or more of the compounds described hereincan be prepared using a pharmaceutically acceptable carrier composed ofmaterials that are considered safe and effective and can be administeredto an individual without causing undesirable biological side effects orunwanted interactions. The carrier is all components present in thepharmaceutical formulation other than the active ingredient oringredients. As generally used herein “carrier” includes, but is notlimited to, diluents, binders, lubricants, disintegrators, fillers, pHmodifying agents, preservatives, antioxidants, solubility enhancers, andcoating compositions.

Carrier also includes all components of the coating composition whichcan include plasticizers, pigments, colorants, stabilizing agents, andglidants. Delayed release, extended release, and/or pulsatile releasedosage formulations can be prepared as described in standard referencessuch as “Pharmaceutical dosage form tablets,” eds. Liberman et. al. (NewYork, Marcel Dekker, Inc., 1989), “Remington—The science and practice ofpharmacy,” 20th ed., Lippincott Williams & Wilkins, Baltimore, Md.,2000, and “Pharmaceutical dosage forms and drug delivery systems,” 6thEdition, Ansel et al., (Media, Pa.: Williams and Wilkins, 1995). Thesereferences provide information on carriers, materials, equipment andprocess for preparing tablets and capsules and delayed release dosageforms of tablets, capsules, and granules.

Examples of suitable coating materials include, but are not limited to,cellulose polymers such as cellulose acetate phthalate, hydroxypropylcellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulosephthalate and hydroxypropyl methylcellulose acetate succinate; polyvinylacetate phthalate, acrylic acid polymers and copolymers, and methacrylicresins that are commercially available under the trade name EUDRAGIT®(Roth Pharma, Westerstadt, Germany), zein, shellac, and polysaccharides.

Additionally, the coating material can contain conventional carrierssuch as plasticizers, pigments, colorants, glidants, stabilizationagents, pore formers and surfactants.

Optional pharmaceutically acceptable excipients present in thedrug-containing tablets, beads, granules or particles include, but arenot limited to, diluents, binders, lubricants, disintegrants, colorants,stabilizers, and surfactants.

Diluents, also referred to as “fillers,” are typically necessary toincrease the bulk of a solid dosage form so that a practical size isprovided for compression of tablets or formation of beads and granules.Suitable diluents include, but are not limited to, dicalcium phosphatedihydrate, calcium sulfate, lactose, sucrose, mannitol, sorbitol,cellulose, microcrystalline cellulose, kaolin, sodium chloride, drystarch, hydrolyzed starches, pregelatinized starch, silicone dioxide,titanium oxide, magnesium aluminum silicate and powdered sugar.

Binders are used to impart cohesive qualities to a solid dosageformulation, and thus ensure that a tablet or bead or granule remainsintact after the formation of the dosage forms. Suitable bindermaterials include, but are not limited to, starch, pregelatinizedstarch, gelatin, sugars (including sucrose, glucose, dextrose, lactoseand sorbitol), polyethylene glycol, waxes, natural and synthetic gumssuch as acacia, tragacanth, sodium alginate, cellulose, includinghydroxypropylmethylcellulose, hydroxypropylcellulose, ethylcellulose,and veegum, and synthetic polymers such as acrylic acid and methacrylicacid copolymers, methacrylic acid copolymers, methyl methacrylatecopolymers, aminoalkyl methacrylate copolymers, polyacrylicacid/polymethacrylic acid and polyvinylpyrrolidone.

Lubricants are used to facilitate tablet manufacture. Examples ofsuitable lubricants include, but are not limited to, magnesium stearate,calcium stearate, stearic acid, glycerol behenate, polyethylene glycol,talc, and mineral oil.

Disintegrants are used to facilitate dosage form disintegration or“breakup” after administration, and generally include, but are notlimited to, starch, sodium starch glycolate, sodium carboxymethylstarch, sodium carboxymethylcellulose, hydroxypropyl cellulose,pregelatinized starch, clays, cellulose, alginine, gums or cross linkedpolymers, such as cross-linked PVP (Polyplasdone XL from GAF ChemicalCorp).

Stabilizers are used to inhibit or retard drug decomposition reactionswhich include, by way of example, oxidative reactions.

Surfactants can be anionic, cationic, amphoteric or nonionic surfaceactive agents. Suitable anionic surfactants include, but are not limitedto, those containing carboxylate, sulfonate and sulfate ions. Examplesof anionic surfactants include sodium, potassium, ammonium of long chainalkyl sulfonates and alkyl aryl sulfonates such as sodium dodecylbenzenesulfonate; dialkyl sodium sulfosuccinates, such as sodium dodecylbenzenesulfonate; dialkyl sodium sulfosuccinates, such as sodiumbis-(2-ethylthioxyl)-sulfosuccinate; and alkyl sulfates such as sodiumlauryl sulfate. Cationic surfactants include, but are not limited to,quaternary ammonium compounds such as benzalkonium chloride,benzethonium chloride, cetrimonium bromide, stearyl dimethylbenzylammonium chloride, polyoxyethylene and coconut amine. Examples ofnonionic surfactants include ethylene glycol monostearate, propyleneglycol myristate, glyceryl monostearate, glyceryl stearate,polyglyceryl-4-oleate, sorbitan acylate, sucrose acylate, PEG-150laurate, PEG-400 monolaurate, polyoxyethylene monolaurate, polysorbates,polyoxyethylene octylphenylether, PEG-1000 cetyl ether, polyoxyethylenetridecyl ether, polypropylene glycol butyl ether, Poloxamer® 401,stearoyl monoisopropanolamide, and polyoxyethylene hydrogenated tallowamide. Examples of amphoteric surfactants include sodiumN-dodecyl-.beta.-alanine, sodium N-lauryl-.beta.-iminodipropionate,myristoamphoacetate, lauryl betaine and lauryl sulfobetaine.

If desired, the tablets, beads, granules, or particles can also containminor amount of nontoxic auxiliary substances such as wetting oremulsifying agents, dyes, pH buffering agents, or preservatives.

The compositions described herein can be formulation for modified orcontrolled release. Examples of controlled release dosage forms includeextended release dosage forms, delayed release dosage forms, pulsatilerelease dosage forms, and combinations thereof.

The extended release formulations are generally prepared as diffusion orosmotic systems, for example, as described in “Remington—The science andpractice of pharmacy” (20th ed., Lippincott Williams & Wilkins,Baltimore, Md., 2000). A diffusion system typically consists of twotypes of devices, a reservoir and a matrix, and is well known anddescribed in the art. The matrix devices are generally prepared bycompressing the drug with a slowly dissolving polymer carrier into atablet form. The three major types of materials used in the preparationof matrix devices are insoluble plastics, hydrophilic polymers, andfatty compounds. Plastic matrices include, but are not limited to,methyl acrylate-methyl methacrylate, polyvinyl chloride, andpolyethylene. Hydrophilic polymers include, but are not limited to,cellulosic polymers such as methyl and ethyl cellulose,hydroxyalkylcelluloses such as hydroxypropyl-cellulose,hydroxypropylmethylcellulose, sodium carboxymethylcellulose, andCarbopol® 934, polyethylene oxides and mixtures thereof. Fatty compoundsinclude, but are not limited to, various waxes such as carnauba wax andglyceryl tristearate and wax-type substances including hydrogenatedcastor oil or hydrogenated vegetable oil, or mixtures thereof.

In certain preferred embodiments, the plastic material is apharmaceutically acceptable acrylic polymer, including but not limitedto, acrylic acid and methacrylic acid copolymers, methyl methacrylate,methyl methacrylate copolymers, ethoxyethyl methacrylates, cyanoethylmethacrylate, aminoalkyl methacrylate copolymer, poly(acrylic acid),poly(methacrylic acid), methacrylic acid alkylamine copolymerpoly(methyl methacrylate), poly(methacrylic acid) (anhydride),polymethacrylate, polyacrylamide, poly(methacrylic acid anhydride), andglycidyl methacrylate copolymers.

In certain preferred embodiments, the acrylic polymer is comprised ofone or more ammonio methacrylate copolymers. Ammonio methacrylatecopolymers are well known in the art, and are described in NF XVII asfully polymerized copolymers of acrylic and methacrylic acid esters witha low content of quaternary ammonium groups.

In one preferred embodiment, the acrylic polymer is an acrylic resinlacquer such as that which is commercially available from Rohm Pharmaunder the tradename Eudragit®. In further preferred embodiments, theacrylic polymer comprises a mixture of two acrylic resin lacquerscommercially available from Rohm Pharma under the tradenames Eudragit®RL30D and Eudragit® RS30D, respectively. Eudragit® RL30D and Eudragit®RS30D are copolymers of acrylic and methacrylic esters with a lowcontent of quaternary ammonium groups, the molar ratio of ammoniumgroups to the remaining neutral (meth)acrylic esters being 1:20 inEudragit® RL30D and 1:40 in Eudragit® RS30D. The mean molecular weightis about 150,000. Edragit® S-100 and Eudragit® L-100 are also preferred.The code designations RL (high permeability) and RS (low permeability)refer to the permeability properties of these agents. Eudragit® RL/RSmixtures are insoluble in water and in digestive fluids. However,multiparticulate systems formed to include the same are swellable andpermeable in aqueous solutions and digestive fluids.

The polymers described above such as Eudragit® RL/RS can be mixedtogether in any desired ratio in order to ultimately obtain asustained-release formulation having a desirable dissolution profile.Desirable sustained-release multiparticulate systems can be obtained,for instance, from 100% Eudragit® RL, 50% Eudragit® RL and 50% Eudragit®RS, and 10% Eudragit® RL and 90% Eudragit® RS. One skilled in the artwill recognize that other acrylic polymers can also be used, such as,for example, Eudragit® L.

Alternatively, extended release formulations can be prepared usingosmotic systems or by applying a semi-permeable coating to the dosageform. In the latter case, the desired drug release profile can beachieved by combining low permeable and high permeable coating materialsin suitable proportion.

The devices with different drug release mechanisms described above canbe combined in a final dosage form comprising single or multiple units.Examples of multiple units include, but are not limited to, multilayertablets and capsules containing tablets, beads, or granules. Animmediate release portion can be added to the extended release system bymeans of either applying an immediate release layer on top of theextended release core using a coating or compression process or in amultiple unit system such as a capsule containing extended and immediaterelease beads.

Extended release tablets containing hydrophilic polymers are prepared bytechniques commonly known in the art such as direct compression, wetgranulation, or dry granulation. Their formulations usually incorporatepolymers, diluents, binders, and lubricants as well as the activepharmaceutical ingredient. The usual diluents include inert powderedsubstances such as starches, powdered cellulose, especially crystallineand microcrystalline cellulose, sugars such as fructose, mannitol andsucrose, grain flours and similar edible powders. Typical diluentsinclude, for example, various types of starch, lactose, mannitol,kaolin, calcium phosphate or sulfate, inorganic salts such as sodiumchloride and powdered sugar. Powdered cellulose derivatives are alsouseful. Typical tablet binders include substances such as starch,gelatin and sugars such as lactose, fructose, and glucose. Natural andsynthetic gums, including acacia, alginates, methylcellulose, andpolyvinylpyrrolidone can also be used. Polyethylene glycol, hydrophilicpolymers, ethylcellulose and waxes can also serve as binders. Alubricant is necessary in a tablet formulation to prevent the tablet andpunches from sticking in the die. The lubricant is chosen from suchslippery solids as talc, magnesium and calcium stearate, stearic acidand hydrogenated vegetable oils.

Extended release tablets containing wax materials are generally preparedusing methods known in the art such as a direct blend method, acongealing method, and an aqueous dispersion method. In the congealingmethod, the drug is mixed with a wax material and either spray-congealedor congealed and screened and processed.

Delayed release formulations are created by coating a solid dosage formwith a polymer film, which is insoluble in the acidic environment of thestomach, and soluble in the neutral environment of the small intestine.

The delayed release dosage units can be prepared, for example, bycoating a drug or a drug-containing composition with a selected coatingmaterial. The drug-containing composition can be, e.g., a tablet forincorporation into a capsule, a tablet for use as an inner core in a“coated core” dosage form, or a plurality of drug-containing beads,particles or granules, for incorporation into either a tablet orcapsule. Preferred coating materials include bioerodible, graduallyhydrolyzable, gradually water-soluble, and/or enzymatically degradablepolymers, and can be conventional “enteric” polymers. Enteric polymers,as will be appreciated by those skilled in the art, become soluble inthe higher pH environment of the lower gastrointestinal tract or slowlyerode as the dosage form passes through the gastrointestinal tract,while enzymatically degradable polymers are degraded by bacterialenzymes present in the lower gastrointestinal tract, particularly in thecolon. Suitable coating materials for effecting delayed release include,but are not limited to, cellulosic polymers such as hydroxypropylcellulose, hydroxyethyl cellulose, hydroxymethyl cellulose,hydroxypropyl methyl cellulose, hydroxypropyl methyl cellulose acetatesuccinate, hydroxypropylmethyl cellulose phthalate, methylcellulose,ethyl cellulose, cellulose acetate, cellulose acetate phthalate,cellulose acetate trimellitate and carboxymethylcellulose sodium;acrylic acid polymers and copolymers, preferably formed from acrylicacid, methacrylic acid, methyl acrylate, ethyl acrylate, methylmethacrylate and/or ethyl methacrylate, and other methacrylic resinsthat are commercially available under the tradename Eudragit® (RohmPharma; Westerstadt, Germany), including Eudragit® L30D-55 and L100-55(soluble at pH 5.5 and above), Eudragit® L-100 (soluble at pH 6.0 andabove), Eudragit® S (soluble at pH 7.0 and above, as a result of ahigher degree of esterification), and Eudragits® NE, RL and RS(water-insoluble polymers having different degrees of permeability andexpandability); vinyl polymers and copolymers such as polyvinylpyrrolidone, vinyl acetate, vinylacetate phthalate, vinylacetatecrotonic acid copolymer, and ethylene-vinyl acetate copolymer;enzymatically degradable polymers such as azo polymers, pectin,chitosan, amylose and guar gum; zein and shellac. Combinations ofdifferent coating materials can also be used. Multi-layer coatings usingdifferent polymers can also be applied.

The preferred coating weights for particular coating materials can bereadily determined by those skilled in the art by evaluating individualrelease profiles for tablets, beads and granules prepared with differentquantities of various coating materials. It is the combination ofmaterials, method and form of application that produce the desiredrelease characteristics, which one can determine only from the clinicalstudies.

The coating composition can include conventional additives, such asplasticizers, pigments, colorants, stabilizing agents, glidants, etc. Aplasticizer is normally present to reduce the fragility of the coating,and will generally represent about 10 wt. % to 50 wt. % relative to thedry weight of the polymer. Examples of typical plasticizers includepolyethylene glycol, propylene glycol, triacetin, dimethyl phthalate,diethyl phthalate, dibutyl phthalate, dibutyl sebacate, triethylcitrate, tributyl citrate, triethyl acetyl citrate, castor oil andacetylated monoglycerides. A stabilizing agent is preferably used tostabilize particles in the dispersion. Typical stabilizing agents arenonionic emulsifiers such as sorbitan esters, polysorbates andpolyvinylpyrrolidone. Glidants are recommended to reduce stickingeffects during film formation and drying, and will generally representapproximately 25 wt. % to 100 wt. % of the polymer weight in the coatingsolution. One effective glidant is talc. Other glidants such asmagnesium stearate and glycerol monostearates can also be used. Pigmentssuch as titanium dioxide can also be used. Small quantities of ananti-foaming agent, such as a silicone (e.g., simethicone), can also beadded to the coating composition.

Alternatively, each dosage unit in the capsule can comprise a pluralityof drug-containing beads, granules or particles. As is known in the art,drug-containing “beads” refer to beads made with drug and one or moreexcipients or polymers. Drug-containing beads can be produced byapplying drug to an inert support, e.g., inert sugar beads coated withdrug or by creating a “core” comprising both drug and one or moreexcipients. As is also known, drug-containing “granules” and “particles”comprise drug particles that can or can not include one or moreadditional excipients or polymers. In contrast to drug-containing beads,granules and particles do not contain an inert support. Granulesgenerally comprise drug particles and require further processing.Generally, particles are smaller than granules, and are not furtherprocessed. Although beads, granules and particles can be formulated toprovide immediate release, beads and granules are generally employed toprovide delayed release.

Terms

As used herein, “alkyl” means a noncyclic straight chain or branched,unsaturated or saturated hydrocarbon such as those containing from 1 to10 carbon atoms, while the term “lower alkyl” or “C₁₋₄alkyl” has thesame meaning as alkyl but contains from 1 to 4 carbon atoms. The term“higher alkyl” has the same meaning as alkyl but contains from 7 to 20carbon atoms. Representative saturated straight chain alkyls includemethyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-septyl, n-octyl,n-nonyl, and the like; while saturated branched alkyls includeisopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, and the like.Unsaturated alkyls contain at least one double or triple bond betweenadjacent carbon atoms (referred to as an “alkenyl” or “alkynyl”,respectively). Representative straight chain and branched alkenylsinclude ethylenyl, propylenyl, 1-butenyl, 2-butenyl, isobutylenyl,1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl,2,3-dimethyl-2-butenyl, and the like; while representative straightchain and branched alkynyls include acetylenyl, propynyl, 1-butynyl,2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl, and the like.

Non-aromatic mono or polycyclic alkyls are referred to herein as“carbocycles” or “carbocyclyl” groups. Representative saturatedcarbocycles include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,and the like; while unsaturated carbocycles include cyclopentenyl andcyclohexenyl, and the like.

“Heterocarbocycles” or heterocarbocyclyl” groups are carbocycles whichcontain from 1 to 4 heteroatoms independently selected from nitrogen,oxygen and sulfur which may be saturated or unsaturated (but notaromatic), monocyclic or polycyclic, and wherein the nitrogen and sulfurheteroatoms may be optionally oxidized, and the nitrogen heteroatom maybe optionally quaternized. Heterocarbocycles include morpholinyl,pyrrolidinonyl, pyrrolidinyl, piperidinyl, hydantoinyl, valerolactamyl,oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl,tetrahydropyridinyl, tetrahydroprimidinyl, tetrahydrothiophenyl,tetrahydrothiopyranyl, tetrahydropyrimidinyl, tetrahydrothiophenyl,tetrahydrothiopyranyl, and the like.

“Aryl” means an aromatic carbocyclic monocyclic or polycyclic ring suchas phenyl or naphthyl. Polycyclic ring systems may, but are not requiredto, contain one or more non-aromatic rings, as long as one of the ringsis aromatic.

As used herein, “heteroaryl” refers an aromatic heterocarbocycle having1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, andcontaining at least 1 carbon atom, including both mono- and polycyclicring systems. Polycyclic ring systems may, but are not required to,contain one or more non-aromatic rings, as long as one of the rings isaromatic. Representative heteroaryls are furyl, benzofuranyl,thiophenyl, benzothiophenyl, pyrrolyl, indolyl, isoindolyl, azaindolyl,pyridyl, quinolinyl, isoquinolinyl, oxazolyl, isooxazolyl, benzoxazolyl,pyrazolyl, imidazolyl, benzimidazolyl, thiazolyl, benzothiazolyl,isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl,cinnolinyl, phthalazinyl, and quinazolinyl. It is contemplated that theuse of the term “heteroaryl” includes N-alkylated derivatives such as a1-methylimidazol-5-yl substituent.

As used herein, “heterocycle” or “heterocyclyl” refers to mono- andpolycyclic ring systems having 1 to 4 heteroatoms selected fromnitrogen, oxygen and sulfur, and containing at least 1 carbon atom. Themono- and polycyclic ring systems may be aromatic, non-aromatic ormixtures of aromatic and non-aromatic rings. Heterocycle includesheterocarbocycles, heteroaryls, and the like.

“Alkylthio” refers to an alkyl group as defined above attached through asulfur bridge. An example of an alkylthio is methylthio, (i.e., —S—CH3).

“Alkoxy” refers to an alkyl group as defined above attached through anoxygen bridge. Examples of alkoxy include, but are not limited to,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, s-butoxy, t-butoxy,n-pentoxy, and s-pentoxy. Preferred alkoxy groups are methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, s-butoxy, t-butoxy.

“Alkylamino” refers an alkyl group as defined above attached through anamino bridge. An example of an alkylamino is methylamino, (i.e.,—NH—CH3).

“Alkanoyl” refers to an alkyl as defined above attached through acarbonyl bride (i.e., —(C═O)alkyl).

“Alkylsulfonyl” refers to an alkyl as defined above attached through asulfonyl bridge (i.e., —S(═O)₂alkyl) such as mesyl and the like, and“Arylsulfonyl” refers to an aryl attached through a sulfonyl bridge(i.e., —S(═O)2aryl).

“Alkylsulfinyl” refers to an alkyl as defined above attached through asulfinyl bridge (i.e. —S(═O)alkyl).

The term “substituted” refers to a molecule wherein at least onehydrogen atom is replaced with a substituent. When substituted, one ormore of the groups are “substituents.” The molecule may be multiplysubstituted. In the case of an oxo substituent (“═O”), two hydrogenatoms are replaced. Example substituents within this context may includehalogen, hydroxy, alkyl, alkoxy, nitro, cyano, oxo, carbocyclyl,carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, —NRaRb, —NRaC(═O)Rb,—NRaC(═O)NRaNRb, —NRaC(═O)ORb, —NRaSO2Rb, —C(═O)Ra, —C(═O)ORa,—C(═O)NRaRb, —OC(═O)NRaRb, —ORa, —SRa, —SORa, —S(═O)2Ra, —OS(═O)2Ra and—S(═O)2ORa. Ra and Rb in this context may be the same or different andindependently hydrogen, halogen hydroxyl, alkyl, alkoxy, alkyl, amino,alkylamino, dialkylamino, carbocyclyl, carbocycloalkyl,heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl.

The term “optionally substituted,” as used herein, means thatsubstitution is optional and therefore it is possible for the designatedatom to be unsubstituted.

As used herein, “salts” refer to derivatives of the disclosed compoundswhere the parent compound is modified making acid or base salts thereof.Examples of salts include, but are not limited to, mineral or organicacid salts of basic residues such as amines, alkylamines, ordialkylamines; alkali or organic salts of acidic residues such ascarboxylic acids; and the like. In preferred embodiment the salts areconventional nontoxic pharmaceutically acceptable salts including thequaternary ammonium salts of the parent compound formed, and non-toxicinorganic or organic acids. Preferred salts include those derived frominorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic,phosphoric, nitric and the like; and the salts prepared from organicacids such as acetic, propionic, succinic, glycolic, stearic, lactic,malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic,phenylacetic, glutamic, benzoic, salicylic, sulfanilic,2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, isethionic, and the like.

“Subject” refers any animal, preferably a human patient, livestock, ordomestic pet.

The term “prodrug” refers to an agent that is converted into abiologically active form in vivo. Prodrugs are often useful because, insome situations, they may be easier to administer than the parentcompound. They may, for instance, be bioavailable by oral administrationwhereas the parent compound is not. The prodrug may also have improvedsolubility in pharmaceutical compositions over the parent drug. Aprodrug may be converted into the parent drug by various mechanisms,including enzymatic processes and metabolic hydrolysis.

As used herein, the terms “prevent” and “preventing” include theprevention of the recurrence, spread or onset. It is not intended thatthe present disclosure be limited to complete prevention. In someembodiments, the onset is delayed, or the severity of the disease isreduced.

As used herein, the terms “treat” and “treating” are not limited to thecase where the subject (e.g. patient) is cured and the disease iseradicated. Rather, embodiments, of the present disclosure alsocontemplate treatment that merely reduces symptoms, and/or delaysdisease progression.

As used herein, the term “combination with” when used to describeadministration with an additional treatment means that the agent may beadministered prior to, together with, or after the additional treatment,or a combination thereof.

As used herein, the term “derivative” refers to a structurally similarcompound that retains sufficient functional attributes of the identifiedanalogue. The derivative may be structurally similar because it islacking one or more atoms, substituted, a salt, in differenthydration/oxidation states, or because one or more atoms within themolecule are switched, such as, but not limited to, replacing a oxygenatom with a sulfur atom or replacing a amino group with a hydroxylgroup. The derivative may be a prodrug. Derivatives may be prepare byany variety of synthetic methods or appropriate adaptations presented insynthetic or organic chemistry text books, such as those provide inMarch's Advanced Organic Chemistry: Reactions, Mechanisms, andStructure, Wiley, 6th Edition (2007) Michael B. Smith or DominoReactions in Organic Synthesis, Wiley (2006) Lutz F. Tietze herebyincorporated by reference.

“Cancer” refers any of various cellular diseases with malignantneoplasms characterized by the proliferation of cells. It is notintended that the diseased cells must actually invade surrounding tissueand metastasize to new body sites. Cancer can involve any tissue of thebody and have many different forms in each body area. Within the contextof certain embodiments, whether “cancer is reduced” can be identified bya variety of diagnostic manners known to one skill in the art including,but not limited to, observation the reduction in size or number of tumormasses or if an increase of apoptosis of cancer cells observed, e.g., ifmore than a 5% increase in apoptosis of cancer cells is observed for asample compound compared to a control without the compound. It can alsobe identified by a change in relevant biomarker or gene expressionprofile, such as PSA for prostate cancer, HER2 for breast cancer, orothers.

EXPERIMENTAL Phosphorylation at Ser 184 Results in Inactivation of theProapoptotic Function of BAX

The growth factor GM-CSF-induced BAX phosphorylation results in amarkedly decreased proapoptotic activity of BAX. Gardai et al., J BiolChem, 2004, 279, 21085-21095. To test whether BAX phosphorylation occursin human lung cancer cells, A549 cells were metabolically labeled andtreated with nicotine (1 μM) for 60 min. Results indicate that nicotinepotently stimulates serine phosphorylation of BAX (FIGS. 1A&B).Intriguingly, nicotine significantly prolongs survival of A549 cellsfollowing cisplatin treatment (FIG. 1C), which may occur in a mechanismlikely, at least in part, through BAX phosphorylation. To test whethernicotine induces BAX phosphorylation at ser184, WT, S184A or S184E cDNAin the pcDNA3 mammalian expression vector was transfected into H157cells. Results indicate that nicotine induces phosphorylation of WT butnot S184A or S184E mutant BAX (FIG. 1D), suggesting that nicotinestimulates BAX phosphorylation exclusively at ser184 site. Importantly,expression of the nonphosphorylatable S184A results in more apoptoticcell death as compared to WT. Nicotine can prolong survival of cellsexpressing WT BAX but not the S184A BAX mutant (FIG. 1E). In contrast,the phosphmimetic S184E BAX exhibits no apoptotic activity. Nicotine hasno additional survival effect in cells expressing the S184E BAX mutant(FIG. 1E). These findings reveal that either nicotine-induced ser184site phosphorylation or genetically mimicking ser184 sitephosphorylation (i.e. S184E) results in abrogation of BAX's proapoptoticfunction.

Effect of Small Molecules that Structurally Target the Ser184 Site ofBAX on Apoptosis of Human Lung Cancer Cells or Primary Normal SmallAirway Epithelial Cells (SAEC).

Both SCLC and NSCLC cells express high levels of endogenous BAX (FIG.2A). In contrast, normal small airway epithelial cells (SAEC) express arelatively low level of endogenous BAX (FIG. 2A). These preliminary datasuggest that BAX may be an ideal therapeutic target in human lungcancers. To test whether small molecules that target BAX at the Ser184site induce apoptosis, H1299, A549 or SAEC cells were treated withvarious small molecules (1 μM) for 48 h. The compound2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol (17) has a potent apoptoticeffect on human lung cancer H1299 or A549 cells as compared to the othersmall molecules tested (FIG. 2B) and a significantly less apoptoticeffect on normal small airway epithelial cells that express relative lowlevel of BAX as compared to H1299 or A549 cells (FIG. 2B).

Compounds Prevent Nicotine-Induced BAX Phosphorylation

A549 cells were metabolically labeled with ³²P-orthophosphoric acid andtreated with nicotine in the absence or presence of2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol, 2 or 3 for 60 min. Treatmentsuppresses nicotine-induced BAX phosphorylation. Functionally, nicotinecan prolong survival of A549 cells following treatment with thetherapeutic drug cisplatin but failed to enhance survival aftertreatment of cells with 2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol,suggesting that inhibition of BAX phosphorylation can bock nicotine'ssurvival activity. The compound,2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol, in combination withcisplatin enhances apoptotic cell death suggesting BAX agonists incombination with chemotherapeutics for treating patients with lungcancer.

To test whether 2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol actuallyworks in vivo, the anti-lung cancer efficacy of was tested using nudemice to produce subcutaneous (s.c.) lung tumor xenografts as described.Five-week-old Nu/Nu nude mice were purchased from Harlan. 5×10⁶ of A549cells in a balanced salt solution were injected into s.c. tissue at theflank region of nude mice. The tumors were allowed to grow to an averagevolume of 225-230 mm³ prior to initiation of therapy as described. Threevarious doses of 2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol (25 mg/kg,50 mg/kg or 75 mg/kg) were administered intraperitoneally (i.p.) to miceeach day (q.d.) for two weeks (n=8 mice). 0.5% DMSO vehicle was used asa control (n=8 mice). Tumor volume was estimated by caliper measurements(V=L×W2/2). Preliminary results show that treatments doses were welltolerated and caused significant regression of established lung cancerxenografts (FIGS. 3A&B). Importantly, doses of 50-75 mg/kg are welltolerated without significant toxicities on liver, kidney and heart.

To test whether 2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol inducesapoptosis in vivo, mice with lung cancer were treated with vehiclecontrol or 50 mg/kg for 24 h. Apoptosis in tumor tissues was analyzed byTUNEL assay. Intriguingly, treatment of lung cancer xenograft mice with2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol resulted in apoptosis intumor tissues (FIG. 3C).

Synthetic Methods 2-Methoxy-3-(2-nitro-fluoren-9-ylidenemethyl)-pyridine(CYD-1-76)

To a solution of 2-nitrofluorene (278 mg, 1.32 mmol) and2-methoxy-3-pyridinecarboxyaldehyde (200 mg, 1.46 mmol) in 15 mL ofmethanol was added KF—Al₂O₃ (190 mg, 1.18 mmol). The resulting mixturewas stirred at 72° C. After 8 hrs, TLC indicated that the startingmaterial was gone. 40 mL of CH₂Cl₂ was added into the reaction mixture.The insoluble solid was filtrated, and the filtrate was concentratedunder vacuum to give a yellow solid, which was recrystallized fromalcohol and CH₂Cl₂ to give 306 mg of CYD-1-76 as a yellow solid. ¹H-NMR(600 MHz, CDCl₃) δ 8.88 (s, 1H), 8.32 (m, 10H), 8.10 (m, 22H), 8.01 (d,1H, J=7.2 Hz), 7.94 (s, 3H), 7.53 (m, 7H), 7.48 (m, 1H), 7.43 (d, 1H,J=7.8 Hz), 7.31 (m, 1H), 7.19 (m, 4H), 3.94 (s, 14H).

9-(2-Methoxy-benzylidene)-2-nitro-9H-fluorene (CYD-1-70)

To a solution of 2-nitrofluorene (1.05 g, 5 mmol) and2-methoxybenzaldehyde (0.816 g, 6 mmol) in 20 mL of methanol was addedKF—Al₂O₃ (0.75 g, 4.5 mmol). The resulting mixture was stirred at 72° C.After 6 hrs, TLC indicated that the starting material was gone. 40 mL ofCH₂Cl₂ was added into the reaction mixture. The insoluble solid wasfiltrated, and the filtrate was concentrated under vacuum to give ayellow solid, which was recrystallized from alcohol and CH₂Cl₂ to give1.2 g of CYD-1-70 as a yellow solid. ¹H-NMR (600 MHz, CDCl₃) δ 8.84 (s,0.36H), 8.26 (m, 1.65H), 8.10 (m, 3.05H), 8.02 (s, 0.57H), 7.55 (m, 4H),7.24 (m, 1.24H), 7.10 (m, 0.91H), 3.84 (s, 3H)

2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol (CYD-1-87)

To a solution of 2-nitrofluorene (250 mg, 1.18 mmol) and salicylaldehyde(159 mg, 1.30 mmol) in 10 mL of methanol was added KF—Al₂O₃ (170 mg,1.06 mmol). The resulting mixture was stirred at 72° C. After 6 hrs, TLCindicated that the starting material was gone. 40 mL of CH₂Cl₂ was addedinto the reaction mixture. The insoluble solid was filtrated, and thefiltrate was concentrated under vacuum to give a yellow solid, which waspurified by silica gel column; eluting with 11% EtOAc in hexane afforded125 mg of CYD-1-87 as a yellow solid. ¹H-NMR (600 MHz, CDCl₃) δ 10.05(br s, 1H), 8.39 (s, 1H), 8.27 (dd, 1H, J=2.4 Hz, 8.4 Hz), 8.16 (d, 1H,J=8.4 Hz), 8.09 (m, 2H), 8.04 (s, 1H), 7.52 (m, 3H), 7.37 (m, 1H), 7.05(d, 1H, J=8.4 Hz), 6.96 (t, 1H, J=7.2 Hz).

2-Methoxy-3-(2-nitro-fluoren-9-ylidenemethyl)-pyridine (CYD-1-76)

To a solution of 2-nitrofluorene (278 mg, 1.32 mmol) and2-methoxy-3-pyridinecarboxyaldehyde (200 mg, 1.46 mmol) in 15 mL ofmethanol was added KF—Al₂O₃ (190 mg, 1.18 mmol). The resulting mixturewas stirred at 72° C. After 8 hrs, TLC indicated that the startingmaterial was gone. 40 mL of CH₂Cl₂ was added into the reaction mixture.The insoluble solid was filtrated, and the filtrate was concentratedunder vacuum to give a yellow solid, which was recrystallized fromalcohol and CH₂Cl₂ to give 306 mg of CYD-1-76 as a yellow solid. ¹H-NMR(600 MHz, CDCl₃) δ 8.88 (s, 1H), 8.32 (m, 10H), 8.10 (m, 22H), 8.01 (d,1H, J=7.2 Hz), 7.94 (s, 3H), 7.53 (m, 7H), 7.48 (m, 1H), 7.43 (d, 1H,J=7.8 Hz), 7.31 (m, 1H), 7.19 (m, 4H), 3.94 (s, 14H).

9-(2-Methoxy-benzylidene)-2-nitro-9H-fluorene (CYD-1-70)

To a solution of 2-nitrofluorene (1.05 g, 5 mmol) and2-methoxybenzaldehyde (0.816 g, 6 mmol) in 20 mL of methanol was addedKF—Al₂O₃ (0.75 g, 4.5 mmol). The resulting mixture was stirred at 72° C.After 6 hrs, TLC indicated that the starting material was gone. 40 mL ofCH₂Cl₂ was added into the reaction mixture. The insoluble solid wasfiltrated, and the filtrate was concentrated under vacuum to give ayellow solid, which was recrystallized from alcohol and CH₂Cl₂ to give1.2 g of CYD-1-70 as a yellow solid. ¹H-NMR (600 MHz, CDCl₃) δ 8.84 (s,0.36H), 8.26 (m, 1.65H), 8.10 (m, 3.05H), 8.02 (s, 0.57H), 7.55 (m, 4H),7.24 (m, 1.24H), 7.10 (m, 0.91H), 3.84 (s, 3H). ¹³C-NMR (150 MHz, CDCl₃)δ 157.7, 147.4, 146.6, 146.5, 144.2, 140.8, 140.0, 138.6, 137.9, 136.8,136.6, 134.2, 134.0, 131.5, 131.2, 131.1, 130.8, 129.6, 129.5, 129.2(2C), 128.9, 128.4, 124.5, 124.3, 124.2, 124.1, 123.9, 122.2, 121.9,121.6, 121.1, 120.9, 120.8 (2C), 119.1, 116.6 (2C), 112.1, 112.0, 55.9(2C).

3-((2-Nitro-9H-fluoren-9-ylidene)methyl)pyridin-2(1H)-one (CYD-1-93)

To a solution of 2-nitrofluorene (326 mg, 1.54 mmol) and2-oxo-1,2-dihydro-pyridine-3-carbaldehyde (19 mg, 1.54 mmol) in 10 mL ofmethanol was added KF—Al₂O₃ (224 mg, 1.38 mmol). The resulting mixturewas stirred at 85° C. After 24 hrs, TLC indicated that a new product wasproduced and lots of starting material was still remained. 40 mL ofCH₂Cl₂ was added into the reaction mixture. The insoluble solid wasfiltrated, and the filtrate was concentrated under vacuum to give ayellow solid, which was purified by silica gel column; eluting with 60%EtOAc in hexane afforded 26 mg of CYD-1-93 as a yellow solid. ¹H-NMR(600 MHz, d₆-DMSO) δ 12.18 (br s, 2H), 8.77 (d, 1H, J=1.8 Hz), 8.52 (d,1H, J=1.8 Hz), 8.28 (m, 2H), 8.15 (m, 2H), 8.10 (d, 1H, J=7.8 Hz), 8.06(m, 2H), 7.97 (m, 2H), 7.86 (m, 2H), 7.80 (s, 1H), 7.63 (m, 1H), 7.59(m, 1H), 7.49 (m, 3H), 7.37 (m, 1H), 6.40 (m, 2H). ¹³C-NMR (150 MHz,CDCl₃) δ 161.2, 161.1, 146.9, 146.2, 146.1, 143.6, 142.1, 141.7, 140.5,139.6, 138.3, 137.3, 136.1, 136.0, 133.6, 133.4, 129.3, 129.1, 129.0,128.8, 127.9, 127.4, 126.1, 125.9, 124.0, 123.9, 123.5, 121.9, 121.5,121.1, 120.7, 120.5, 119.0, 105.1, 105.0.

9-(2-Methoxy-benzylidene)-9H-fluoren-2-ylamine (CYD-1-96)

To a solution of CYD-1-70 (100 mg, 0.304 mmol) in 10 mL of THF was added0.4 mL of sat. NH₄Cl and 0.4 mL of H₂O. The resulting mixture was cooledto 0° C. in an ice-water bath. Then 236 mg of Zinc dust was added intoit at 0° C. The reaction was stirred at rt for 2 hrs. TLC indicated thatthe starting material was gone. The Zinc solid was filtrated, and thefiltrate was concentrated under vacuum to give a yellow residue, whichwas purified by silica gel column; eluting with 33% EtOAc in hexaneafforded 90 mg of CYD-1-96 (100%) as yellow oil. One isomer: ¹H-NMR (600MHz, CDCl₃) δ 7.71 (d, 1H, J=7.2 Hz), 7.62 (m, 2H), 7.51 (m, 1H), 7.42(m, 1H), 7.33 (m, 1H), 7.26 (m, 1H), 7.18 (m, 1H), 6.94 (m, 3H), 6.56(m, 1H), 3.80 (s, 3H); Another isomer: 7.62 (m, 1H), 7.54 (s, 1H), 7.51(m, 2H), 7.42 (m, 1H), 7.33 (m, 1H), 7.18 (m, 2H), 7.04 (d, 1H, J=1.8Hz), 6.94 (m, 2H), 6.64 (m, 1H), 3.80 (s, 3H). ¹³C-NMR (150 MHz, CDCl₃)δ 157.7, 157.6, 146.0, 145.5, 141.9, 141.4, 139.7, 139.1, 138.4, 136.3,136.3, 136.2, 132.7, 131.2, 131.2, 130.6, 129.8, 129.8, 128.3, 128.0,125.6, 125.6, 125.3, 124.9, 124.1, 123.5, 123.3, 120.4, 120.3, 120.3,120.3, 120.2, 118.4, 118.2, 115.5, 115.3, 111.1, 110.8, 110.8, 107.2,55.5, 55.5.

4-{2-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-morpholine(CYD-1-95)

To a solution of CYD-1-94 (80 mg, 0.25 mmol) in 8 mL of THF was addedPPh₃ (117.9 mg, 0.45 mmol) and 2-morpholin-4-yl-ethanol (59 mg, 0.45mmol). Then DIAD (91 mg, 0.45 mmol) was added into the resultingmixture. The reaction mixture was stirred at rt for 3 hrs. After that,TLC showed CYD-1-94 was gone. The solvent was removed under vacuum togive a yellow residue, which was purified by silica gel column; elutingwith EtOAc afforded 87 mg of CYD-1-93 as yellow oil. ¹H-NMR (600 MHz,CDCl₃) δ 8.63 (d, 1H, J=1.8 Hz), 8.41 (d, 1H, J=1.8 Hz), 8.25 (dd, 1H,J=1.8 Hz, 8.4 Hz), 8.19 (dd, 1H, J=1.8 Hz, 7.8 Hz), 7.85 (m, 3H), 7.80(m, 4H), 7.67 (d, 1H, J=7.8 Hz), 7.63 (d, 1H, J=7.2 Hz), 7.58 (d, 1H,J=6.6 Hz), 7.41 (m, 5H), 7.19 (m, 1H), 7.10 (m, 1H), 7.04 (m, 3H), 4.19(m, 4H), 3.58 (m, 8H), 2.76 (t, 2H, J=6.0 Hz), 2.71 (t, 2H, J=6.0 Hz),2.48 (s, 8H). ¹³C-NMR (150 MHz, CDCl₃) δ 156.9, 156.8, 147.0, 146.3,144.3, 141.0, 140.2, 138.7, 138.2, 137.1, 136.7, 134.4, 134.2, 130.8,130.8, 130.8, 130.5, 128.8, 128.7, 128.5, 128.4, 127.0, 124.8, 124.5,124.4, 123.6, 123.3, 120.9, 120.8, 120.8, 120.6, 120.5, 119.7, 119.5,119.4, 115.8, 112.2, 112.1, 66.8, 66.7, 66.7, 66.7, 57.4 (2C), 54.0(6C).

1-{2-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-piperazine(CYD-2-7-1)

To a solution of CYD-1-94 (120 mg, 0.38 mmol) in 8 mL of THF was addedPPh₃ (179 mg, 0.68 mmol) and 4-(2-hydroxy-ethyl)-piperazine-1-carboxylicacid tert-butyl ester (157 mg, 0.68 mmol). Then DIAD (138 mg, 0.68 mmol)was added into the resulting mixture. The reaction mixture was stirredat rt for 3 hrs. After that, TLC showed CYD-1-94 was gone. The solventwas removed under vacuum to give a yellow residue, which was purified bysilica gel column; eluting with 50% EtOAc in hexane afforded 196 mg ofCYD-2-7 as yellow oil. CYD-2-7 (196 mg, 0.37 mmol) was dissolved in 4 mLof CH₂Cl₂, and then 1 mL of TFA was added into it at 0° C. The resultingmixture was stirred at rt for 4 hrs. After that, TLC showed that CYD-2-7disappeared. The reaction mixture was washed with sat. NaHCO₃, andconcentrated under vacuum to give an oil residue, which was purified bysilica gel column; eluting with CH₂Cl₂/MeOH/Et₃N=10:1:0.3 afforded 160mg of CYD-2-7-1 as yellow oil. ¹H-NMR (600 MHz, CDCl₃) δ 8.57 (s, 1H),8.37 (s, 1H), 8.13 (m, 2H), 7.82 (m, 3H), 7.70 (m, 4H), 7.65 (d, 1H,J=7.8 Hz), 7.59 (d, 1H, J=7.2 Hz), 7.55 (d, 1H, J=7.2 Hz), 7.41 (m, 4H),7.33 (t, 1H, J=7.2 Hz), 7.16 (t, 1H, J=7.2 Hz), 7.07 (m, 1H), 7.02 (m,3H), 4.82 (br s, 2H), 4.17 (m, 4H), 2.82 (m, 8H), 2.76 (t, 2H, J=6.0Hz), 2.71 (t, 2H, J=5.4 Hz), 2.53 (m, 8H). ¹³C-NMR (150 MHz, CDCl₃) δ156.8 (2C), 146.8, 146.5, 146.2, 144.1, 140.9, 140.1, 138.5, 138.0,136.9, 136.6, 134.2, 134.0, 130.9, 130.8 (2C), 130.6, 128.8, 128.7,128.5, 128.3, 127.0 (2C), 124.6, 124.3, 123.5 (2C), 123.2, 120.9, 120.8,120.7, 120.5 (2C), 119.6 (2C), 119.4 (2C), 115.7, 112.1, 112.0, 66.6,66.5, 57.3, 57.2, 53.6 (2C), 45.2 (6C).

2-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethanol (CYD-2-1)

To a solution of CYD-1-94 (120 mg, 0.38 mmol) in 8 mL of DMF was addedNaH (12 mg, 0.49 mmol). The color of mixture turned into dark red. After5 min, 2-bromoethanol (142 mg, 1.14 mmol) was added into the resultingmixture. The reaction was stirred at 60° C. for 24 hrs. After that, TLCshowed most of CYD-1-94 was gone. The DMF solvent was removed at 60° C.under vacuum to give a yellow oil residue, which was purified by silicagel column; eluting with 80% EtOAc in hexane afforded 86 mg of CYD-2-1as a yellow solid. ¹H-NMR (600 MHz, CDCl₃) δ 8.84 (d, 1H, J=1.2 Hz),8.34 (d, 1H, J=1.8 Hz), 8.31 (dd, 1H, J=1.8 Hz, 8.4 Hz), 8.27 (dd, 1H,J=1.8 Hz, 7.8 Hz), 8.17 (m, 3H), 8.09 (m, 4H), 7.62 (m, 3H), 7.53 (m,3H), 7.48 (m, 2H), 7.29 (t, 1H, J=7.8 Hz), 7.23 (m, 2H), 7.10 (m, 2H),4.86 (t, 1H, J=5.4 Hz), 4.82 (t, 1H, J=5.4 Hz), 4.11 (m, 4H), 3.66 (m,4H).

2-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethylamine (CYD-2-11)

To a solution of CYD-1-94 (150 mg, 0.47 mol) in 8 mL of THF was addedPPh₃ (224 mg, 0.85 mmol) and (2-hydroxy-ethyl)-carbamic acid tert-butylester (138 mg, 0.85 mmol). Then DIAD (173 mg, 0.85 mmol) was added intothe resulting mixture. The reaction mixture was stirred at rt for 4 hrs.After that, TLC showed CYD-1-94 was gone. The solvent was removed undervacuum to give a yellow residue, which was purified by silica gelcolumn; eluting with 80% EtOAc in hexane afforded 160 mg of CYD-2-10 asyellow oil. CYD-2-10 (160 mg, 0.34 mmol) was dissolved in 4 mL ofCH₂Cl₂, and then 1 mL of TFA was added into it at 0° C. The resultingmixture was stirred at rt for 4 hrs. After that, TLC showed thatCYD-2-10 disappeared. The reaction mixture was washed with sat. NaHCO₃,and concentrated under vacuum to give an oil residue, which was purifiedby silica gel column; eluting with CH₂Cl₂/MeOH/Et₃N=10:1:0.3 afforded125 mg of CYD-2-7-1 as yellow oil. ¹H-NMR (600 MHz, CDCl₃) δ 8.66 (d,2H, J=1.8 Hz), 8.43 (d, 2H, J=1.8 Hz), 8.26 (dd, 2H, J=1.8 Hz, 8.4 Hz),8.20 (dd, 1H, J=1.8 Hz, 9.0 Hz), 7.91 (m, 2H), 7.86 (s, 2H), 7.80 (m,6H), 7.67 (d, 2H, J=7.8 Hz), 7.63 (d, 2H, J=7.8 Hz), 7.60 (d, 1H, J=7.8Hz), 7.44 (m, 7H), 7.21 (t, 2H, J=7.8 Hz), 7.07 (m, 6H), 4.10 (m, 6H),3.04 (br s, 6H), 2.66 (m, 6H). ¹³C-NMR (150 MHz, CDCl₃) δ 156.6, 146.9,146.5, 146.3, 144.3, 140.9, 140.1, 138.6, 138.1, 137.0, 136.6, 134.4(2C), 131.0 (2C), 130.9, 130.6, 128.9, 128.8, 128.5, 128.4, 126.7 (2C),124.6, 124.4, 124.3, 123.6, 123.3, 120.9 (2C), 120.8, 120.7, 120.6,119.6, 119.5 (2C), 115.9, 112.1, 112.0, 69.9, 40.8.

4-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-piperidine (CYD-2-13)

To a solution of CYD-1-94 (155 mg, 0.49 mol) in 8 mL of THF was addedPPh₃ (232 mg, 0.88 mmol) and 4-hydroxy-piperidine-1-carboxylic acidtert-butyl ester (178 mg, 0.88 mmol). Then DIAD (178 mg, 0.88 mmol) wasadded into the resulting mixture. The reaction mixture was stirred at rtfor 4 hrs. After that, TLC showed CYD-1-94 was gone. The solvent wasremoved under vacuum to give a yellow residue, which was purified bysilica gel column; eluting with 25% EtOAc in hexane afforded 210 mg ofCYD-2-12 as yellow oil. CYD-2-12 (210 mg, 0.42 mmol) was dissolved in 4mL of CH₂Cl₂, and then 1 mL of TFA was added into it at 0° C. Theresulting mixture was stirred at rt for 4 hrs. After that, TLC showedthat CYD-2-12 disappeared. The reaction mixture was washed with sat.NaHCO₃, and concentrated under vacuum to give an oil residue, which waspurified by silica gel column; eluting with CH₂Cl₂/MeOH/Et₃N=15:1:0.3afforded 140 mg of CYD-2-13 as yellow oil. ¹H-NMR (600 MHz, CDCl₃) δ8.57 (s, 1H), 8.40 (s, 1H), 8.18 (d, 1H, J=8.4 Hz), 8.12 (d, 1H, J=7.8Hz), 7.86 (m, 2H), 7.80 (s, 1H), 7.72 (m, 4H), 7.59 (m, 3H), 7.41 (m,4H), 7.33 (t, 1H, J=7.2 Hz), 7.16 (m, 1H), 7.06 (t, 1H, J=7.2 Hz), 7.02(m, 3H), 5.79 (br s, 2H), 4.53 (m, 2H), 3.06 (m, 4H), 2.81 (m, 4H), 2.03(m, 4H), 1.79 (m, 4H). ¹³C-NMR (150 MHz, CDCl₃) δ 155.3, 155.2, 147.0,146.6, 146.4, 144.3, 140.9, 140.1, 138.7, 138.1, 136.9, 136.6, 134.4(2C), 131.3, 131.2, 130.7, 130.4, 128.9, 128.8, 128.5, 128.4, 126.8,125.9, 125.5, 124.4, 123.6, 123.3, 121.1, 120.9 (2C), 120.8, 120.6,119.6, 119.5, 119.4, 115.7, 114.1 (2C), 72.3, 71.8, 42.2 (2C), 42.0(2C), 30.3 (2C), 30.0 (2C).

1-(4-Fluoro-benzenesulfonyl)-4-{2-[2-(2-nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-piperazine(CYD-2-18)

To a solution of CYD-2-7-1 (120 mg, 0.28 mmol) in 8 mL of CH₂Cl₂ wasadded Et₃N (56.8 mg, 0.56 mmol) and 4-fluoro-benzenesulfonyl chloride(65 mg, 0.33 mmol). The resulting mixture was stirred at rt for 2 hrs.After that, TLC showed that CYD-2-7-1 was gone. The reaction mixture waswashed with water, and dried with anhydrous Na₂SO₄. The solvent wasremoved under vacuum to give a yellow oil residue, which was purified bysilica gel column; eluting with 50% EtOAc in hexane afforded 101 mg ofCYD-2-18 as a yellow solid. ¹H-NMR (600 MHz, CDCl₃) δ 8.56 (d, 1H, J=1.2Hz), 8.17 (dd, 1H, J=1.2 Hz, 7.8 Hz), 7.78 (s, 1H), 7.76 (d, 1H, J=7.2Hz), 7.71 (d, 1H, J=8.4 Hz), 7.62 (m, 2H), 7.56 (d, 2H, J=7.8 Hz), 7.36(m, 2H), 7.14 (m, 1H), 7.03 (m, 3H), 6.96 (m, 1H). 4.10 (m, 2H), 2.83(s, 4H), 2.73 (t, 2H, J=5.4 Hz), 2.51 (m, 4H); ¹³C-NMR (150 MHz, CDCl₃)δ 166.0, 164.3, 156.7, 156.6, 147.1, 146.7, 146.3, 144.2, 140.9, 140.1,138.7, 138.1, 137.1, 136.7, 134.5 (2C), 134.4, 131.6 (2C), 130.8 (2C),130.5, 130.3 (2C), 128.9, 128.8, 128.6, 128.5, 126.8, 124.8, 124.5 (2C),124.4, 123.6 (2C), 123.4, 121.0 (2C), 120.9, 120.8, 120.4, 119.7 (2C),119.6, 119.5, 116.2 (2C), 116.0 (2C), 115.7, 112.2, 112.1, 66.9, 66.7,56.5 (2C), 52.5 (2C), 45.8 (2C).

1-(4-{2-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-piperazin-1-yl)-ethanone(CYD-2-17)

To a solution of CYD-2-7-1 (130 mg, 0.30 mmol) in 8 mL of CH₂Cl₂ wasadded Et₃N (61 mg, 0.60 mmol) and acetyl chloride (28 mg, 0.36 mmol).The resulting mixture was stirred at rt for 2 hrs. After that, TLCshowed that CYD-2-7-1 was gone. The reaction mixture was washed withwater, and dried with anhydrous Na₂SO₄. The solvent was removed undervacuum to give a yellow oil residue, which was purified by silica gelcolumn; eluting with CH₂Cl₂/MeOH=15:1 afforded 108 mg of CYD-2-17 as ayellow oil (75%). ¹H-NMR (600 MHz, CDCl₃) δ 8.61 (s, 1H), 8.37 (s, 1H),8.22 (m, 1H), 8.18 (m, 1H), 7.84 (s, 3H), 7.78 (m, 4H), 7.66 (d, 1H,J=7.8 Hz), 7.62 (d, 1H, J=7.2 Hz), 7.57 (d, 1H, J=7.8 Hz), 7.44 (m, 4H),7.37 (m, 1H), 7.19 (m, 1H), 7.09 (m, 1H), 7.03 (m, 3H), 4.18 (m, 4H),3.49 (s, 4H), 3.29 (d, 2H, J=4.8 Hz), 3.24 (d, 2H, J=4.2 Hz), 2.79 (m,2H), 2.72 (m, 2H), 2.50 (d, 2H, J=4.2 Hz), 2.43 (m, 6H), 1.97 (s, 3H),1.96 (s, 3H); ¹³C-NMR (150 MHz, CDCl₃) δ 168.8, 168.7, 156.8, 156.6,146.9, 146.6, 146.2, 144.2, 140.9, 140.1, 138.6, 138.1, 137.0, 136.7,134.4, 134.2, 130.8 (2C), 130.6, 128.9, 128.8, 128.6, 128.5, 127.0 (2C),124.7, 124.4, 123.6, 123.3 (2C), 120.9 (3C), 119.7 (2C), 119.5 (2C),115.7, 112.2, 112.0, 66.8, 66.7, 56.8 (2C), 53.6, 53.5, 53.1, 53.0,46.1, 46.0, 41.2 (2C), 21.1 (2C)

1-Cyclopropanesulfonyl-4-{2-[2-(2-nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-piperazine(CYD-2-16)

To a solution of CYD-2-7-1 (140 mg, 0.32 mmol) in 8 mL of CH₂Cl₂ wasadded Et₃N (66 mg, 0.65 mmol) and cyclopropanesulfonyl chloride (55 mg,0.39 mmol). The resulting mixture was stirred at rt for 4 hrs. Afterthat, TLC showed that CYD-2-7-1 was gone. The reaction mixture waswashed with water, and dried with anhydrous Na₂SO₄. The solvent wasremoved under vacuum to give a yellow oil residue, which was purified bysilica gel column; eluting with EtOAc/MeOH=40:1 afforded 109 mg ofCYD-2-16 as a yellow solid. ¹H-NMR (600 MHz, CDCl₃) δ 8.64 (d, 1H, J=1.2Hz), 8.38 (d, 1H, J=1.8 Hz), 8.26 (dd, 1H, J=1.8 Hz, 8.4 Hz), 8.20 (dd,1H, J=1.8 Hz, 7.8 Hz), 7.87 (m, 3H), 7.81 (m, 4H), 7.66 (m, 2H), 7.58(d, 1H, J=7.2 Hz), 7.43 (m, 5H), 7.22 (m, 1H), 7.11 (m, 1H), 7.05 (m,3H), 4.19 (m, 4H), 3.17 (m, 8H), 2.80 (t, 2H, J=6.0 Hz), 2.76 (t, 2H,J=5.4 Hz), 2.57 (m, 8H), 2.17 (m, 2H), 1.09 (m, 4H), 0.91 (m, 4H).¹³C-NMR (150 MHz, CDCl₃) δ 156.8, 156.7, 147.1, 146.7, 146.3, 144.3,140.9, 140.1, 138.7, 138.2, 137.1, 136.7, 134.5, 134.3, 130.8 (3C),130.5, 128.9, 128.8, 128.5, 126.9, 124.9, 124.5, 124.4, 123.6, 123.4,121.0 (2C), 120.9, 120.8, 120.4, 119.7 (2C), 119.6, 119.5, 115.7, 112.3,112.2, 66.8, 66.6, 56.9, 56.6, 52.9 (6C), 45.9 (2C), 25.3, 25.2, 4.2(2C), 4.1 (2C).

1-Methanesulfonyl-4-{2-[2-(2-nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-piperazine(CYD-2-26)

To a solution of CYD-2-7-1 (150 mg, 0.35 mmol) in 8 mL of CH₂Cl₂ wasadded Et₃N (70 mg, 0.70 mmol) and methanesulfonyl chloride (48 mg, 0.42mmol). The resulting mixture was stirred at rt for 4 hrs. After that,TLC showed that CYD-2-7-1 was gone. The reaction mixture was washed withwater, and dried with anhydrous Na₂SO₄. The solvent was removed undervacuum to give a yellow oil residue, which was purified by silica gelcolumn; eluting with EtOAc/MeOH=50:1 afforded 130 mg of CYD-2-26 as ayellow solid. ¹H-NMR (600 MHz, CDCl₃) δ 8.58 (d, 1H, J=2.4 Hz), 8.33 (d,1H, J=2.4 Hz), 8.21 (dd, 1H, J=1.8 Hz, 8.4 Hz), 8.15 (dd, 1H, J=1.8 Hz,7.8 Hz), 7.82 (m, 3H), 7.76 (m, 4H), 7.64 (d, 1H, J=7.8 Hz), 7.60 (d,1H, J=7.8 Hz), 7.54 (d, 1H, J=7.2 Hz), 7.39 (m, 5H), 7.17 (t, 1H, J=7.2Hz), 7.08 (m, 1H), 7.01 (m, 3H), 4.14 (m, 4H), 3.06 (m, 4H), 3.02 (m,4H), 2.77 (t, 2H, J=5.4 Hz), 2.72 (t, 2H, J=5.4 Hz), 2.65 (s, 3H), 2.63(s, 3H), 2.53 (m, 8H). ¹³C-NMR (150 MHz, CDCl₃) δ 156.8, 156.7, 147.0,146.7, 146.3, 144.3, 140.9, 140.1, 138.7, 138.1, 137.1, 136.7, 134.5,134.3, 130.9, 130.8 (2C), 130.6, 128.9, 128.8, 128.6, 126.9, 124.9,124.5, 124.4, 123.6 (2C), 123.4, 121.0, 120.9, 120.8, 120.5, 119.7 (2C),119.6, 119.5, 115.7, 112.3 (2C), 66.7, 66.5, 56.5 (2C), 52.7 (3C), 52.6(3C), 45.7 (2C), 34.1, 34.0.

The dimmer of cyclopropanesulfonic acid (9H-fluoren-2-yl)-amide(CYD-2-31)

To a solution of cyclopropanesulfonic acid (9H-fluoren-2-yl)-amide (300mg, 1.05 mmol) in 8 mL of THF was added salicylaldehyde (128 mg, 1.05mmol), 40% NaOH (50 mg, 1.26 mmol) and Bu₄N⁺Cl⁻ (29 mg, 0.10 mmol), Theresulting mixture was stirred at 65° C. for 48 hrs. After that, TLCshowed that a new product was produce, and about half of the startingmaterial was still remained. The reaction mixture was acidized with 10%HCl, and extracted with EtOAc for 3 times. The combined organic phasewas concentrated under vacuum to give a yellow solid residue, which waspurified by silica gel column; eluting with EtOAc/hexane=1:8 afforded 90mg of CYD-2-31 as a yellow solid. ¹H-NMR (600 MHz, d₆-DMSO) δ10.48 (s,2H), 8.12 (s, 2H), 7.98 (m, 6H), 7.76 (d, 2H J=7.8 Hz), 7.72 (d, 2H,J=7.2 Hz), 7.57 (m, 10H), 7.32 (m, 2H). ¹³C-NMR (150 MHz, CDCl₃) δ 166.1(2C), 144.5 (2C), 140.8 (2C), 139.1 (2C), 135.8 (2C), 134.9 (2C), 134.3(2C), 133.9 (2C), 132.2 (2C), 129.1 (2C), 128.8 (4C), 128.1 (4C), 126.2(2C), 124.3 (2C), 121.9 (2C), 121.1 (2C), 116.2 (4C).

The dimmer of N-(9H-fluoren-2-yl)-3-nitro-benzenesulfonamide (CYD-2-38)

To a solution of N,N-di (3-nitro-benzenesulfonamide)-9H-fluoren-2-yl(800 mg, 1.45 mmol) in 20 mL of THF was added salicylaldehyde (212 mg,1.74 mmol), 40% NaOH (75 mg, 1.89 mmol) and Bu₄N⁺Cl⁻ (40 mg, 0.14 mmol).The resulting mixture was stirred at 65° C. for 48 hrs. After that, TLCshowed that a new product was produce, and about half of the startingmaterial was still remained. The reaction mixture was acidized with 10%HCl, and extracted with EtOAc for 3 times. The combined organic phasewas concentrated under vacuum to give a yellow solid residue, which waspurified by silica gel column; eluting with CH₂Cl₂ afforded 130 mg ofCYD-2-38 as a yellow solid. ¹H-NMR (600 MHz, CDCl₃+CD₃OD) δ 8.67 (s,2H), 8.37 (d, 2H, J=8.4 Hz), 8.13 (d, 2H, J=8.4 Hz), 7.68 (t, 2H, J=7.8Hz), 7.57 (d, 2H, J=7.8 Hz), 7.41 (m, 10H), 7.25 (m, 1H). ¹³C-NMR (150MHz, CDCl₃) δ 148.1 (2C), 143.8 (2C), 141.3 (2C), 141.1 (2C), 137.4(2C), 135.1 (2C), 133.8 (2C), 132.5 (2C), 130.4 (2C), 128.8 (2C), 127.2(2C), 127.0 (2C), 124.3 (2C), 122.1 (2C), 121.2 (4C), 120.2 (2C), 117.1(4C).

The dimmer of cyclopropanesulfonic acid (9H-fluoren-2-yl)-amide(CYD-2-36)

To a solution of cyclopropanesulfonic acid (9H-fluoren-2-yl)-amide (250mg, 0.87 mmol) in 10 mL of THF was added salicylaldehyde (117 mg, 0.96mmol), 40% NaOH (42 mg, 1.05 mmol) and Bu₄N⁺Cl⁻ (20 mg, 0.07 mmol). Theresulting mixture was stirred at 65° C. for 48 hrs. After that, TLCshowed that a new product was produce, and about half of the startingmaterial was still remained. The reaction mixture was acidized with 10%HCl, and extracted with EtOAc for 3 times. The combined organic phasewas concentrated under vacuum to give a yellow solid residue, which waspurified by silica gel column; eluting with CH₂Cl₂ afforded 130 mg ofCYD-2-36 as a yellow solid. ¹H-NMR (600 MHz, CDCl₃+CD₃OD) δ 7.61 (d, 2H,J=6.6 Hz), 7.48 (m, 10H), 7.27 (m, 2H), 3.42 (br s, 1H), 2.53 (m, 2H),1.18 (m, 4H), 0.99 (m, 4H). ¹³C-NMR (150 MHz, CDCl₃) δ 144.2 (2C), 140.6(2C), 138.7 (2C), 135.2 (4C), 134.0 (2C), 128.7 (2C), 127.0 (2C), 124.4(2C), 121.2 (2C), 120.2 (4C), 117.2 (2C), 29.9 (2C), 5.4 (4C).

1-(4-Chloro-benzyl)-3-(2-nitro-fluoren-9-ylidenemethyl)-1H-indole(CYD-2-21)

To a solution of 2-nitrofluorene (250 mg, 1.18 mmol) in 20 mL ofmethanol was added 1-(4-Chloro-benzyl)-1H-indole-3-carbaldehyde (382 mg,1.42 mmol) and KF—Al₂O₃ (189 mg, 1.18 mmol). The resulting mixture wasstirred at 85° C. for 18 hrs. After that, TLC showed that2-nitrofluorene was gone, and many solids were suspended in MeOH. 260 mgof CYD-2-21 was obtained as a yellow solid after filtration andrecrystallization from CH₂Cl₂. One isomer: ¹H-NMR (600 MHz, d₆-DMSO) δ8.96 (s, 1H), 8.41 (s, 1H), 8.27 (m, 2H), 8.18 (m, 3H), 8.09 (d, 1H,J=7.2 Hz), 7.78 (d, 1H, J=7.8 Hz), 7.62 (d, 1H, J=7.8 Hz), 7.47 (m, 5H),7.28 (t, 1H, J=7.2 Hz), 7.20 (t, 1H, J=7.2 Hz), 5.57 (s, 2H). Anotherisomer: ¹H-NMR (600 MHz, d₆-DMSO) δ 9.01 (s, 1H), 8.39 (s, 1H), 8.33 (s,1H), 8.19 (m, 4H), 7.86 (d, 1H, J=8.4 Hz), 7.78 (d, 1H, J=7.8 Hz), 7.62(d, 1H, J=7.8 Hz), 7.47 (m, 5H), 7.28 (t, 1H, J=7.2 Hz), 7.20 (t, 1H,J=7.2 Hz), 5.59 (s, 2H). ¹³C-NMR (150 MHz, d₆-DMSO) δ 147.1, 146.6,145.6, 142.9, 141.9, 141.0, 138.2, 138.0, 137.0, 136.8, 136.7, 136.4(2C), 135.6, 132.7, 132.2, 132.1, 130.2, 129.9, 129.8, 129.7, 129.2,129.1, 129.0 (2C), 128.4, 128.1 (2C), 127.9, 124.5, 124.1, 123.8, 123.2(2C), 123.1, 122.4, 122.0, 121.7, 121.2, 121.0 (2C), 120.8, 120.6 (2C),120.2, 118.8, 118.7, 116.2 (2C), 111.5, 111.4, 111.3 (2C), 49.4, 49.2.

2-[3-(2-Nitro-fluoren-9-ylidenemethyl)-pyridin-2-yloxy]-ethylamine(CYD-4-61)

To a solution of 2-fluoro-pyridine-3-carbaldehyde (500 mg, 3.995 mmol)and (2-hydroxy-ethyl)-carbamic acid tert-butyl ester (1287 mg, 7.99mmol) in 20 mL of DMF was added Na₂CO₃ (847 mg, 7.99 mmol). Theresulting mixture was stirred at 80° C. for 5 hrs and the reactionprogress was monitored by TLC analysis. The reaction mixture was thenwashed with brine, and concentrated under vacuum to give an oil residue,which was purified by silica gel column; eluting with EtOAc/hexane=1:2to afford 600 mg of CYD-5-75 in 60% yield as colorless gel. To asolution of 2-nitrofluorene (244 mg, 1.15 mmol) and CYD-5-75 (220 mg,0.82 mmol) in 20 mL of methanol was added KF—Al₂O₃ (184 mg, 1.15 mmol).The resulting mixture was stirred at 72° C. After 6 hrs, TLC indicatedthat the starting material was gone. 40 mL of CH₂Cl₂ was added into thereaction mixture. The insoluble solid was filtrated, and the filtratewas concentrated under vacuum to give a yellow solid, which wasrecrystallized from alcohol and CH₂Cl₂ to give 120 mg of a yellow solid.The yellow solid (180 mg, 0.39 mmol) was dissolved in 4 mL of CH₂Cl₂,and then 1 mL of TFA was added into it at 0° C. The resulting mixturewas stirred at rt for 4 hrs. The reaction mixture was washed with sat.NaHCO₃ (aq.), and concentrated under vacuum to give an oil residue,which was purified by silica gel column; eluting with CH₂Cl₂/MeOH=20:1to provide 150 mg of CYD-4-61 as yellow solid in 50% yield for twosteps. ¹H-NMR (600 MHz, CDCl₃) δ 8.61 (d, 1H, J=1.8 Hz), 8.40 (d, 1H,J=2.4 Hz), 8.27 (m, 2H), 8.23 (dd, 1H, J=2.4 Hz, 8.4 Hz), 8.18 (dd, 1H,J=1.8 Hz, 7.8 Hz), 7.89 (m, 3H), 7.76 (m, 4H), 7.72 (s, 1H), 7.68 (s,1H), 7.61 (d, 1H, J=7.8 Hz), 7.45 (m, 2H), 7.39 (m, 1H), 7.23 (m, 1H),7.04 (m, 1H), 7.00 (m, 1H), 4.46 (m, 4H), 3.07 (m, 4H), 1.45 (br s, 4H).¹³C-NMR (150 MHz, CDCl₃) δ 161.3, 161.2, 148.0, 147.7, 147.1, 146.6(2C), 144.4, 140.7, 139.9, 139.5, 139.4, 138.9, 137.8, 136.8, 136.6,135.5, 135.4, 129.2, 129.0, 128.9, 128.7, 124.6, 124.5, 124.3, 124.0,123.7, 121.2, 120.9, 119.7, 119.6, 119.4, 118.7, 118.4, 116.7, 116.6,116.1, 68.8, 68.7, 41.3 (2C). HRMS calc. for C₂₁H₁₇N₃O₃ [M+H]⁺ 360.1343;found 360.1351. HPLC purity 99.2%

Using appropriate starting materials and the same or appropriatelymodified protocols, the follow compounds were prepared and tested (SeeFIG. 5B, C). In certain embodiments the compounds are selected from.

or salts thereof.

Assays Suppressing Lung Cancer Growth

To compare sensitivities of the compounds, A549 human lung cancer cellswere treated with increasing concentrations (0, 1, 5, 10, 25 μM) of2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol (CYD-1-87) and derivativesfor 48 h. The surviving cell fraction was determined using thesulforhodamine B (SRB) assay as described in (Vichai & Kirtikara, NatProtoc 1, 1112-1116, 2006 hereby incorporated by reference). Thesulforhodamine B (SRB) assay is used for cell density determination,based on the measurement of cellular protein content. The methoddescribed here has been optimized for the toxicity screening ofcompounds to adherent cells in a 96-well format. After an incubationperiod, cell monolayers are fixed with 10% (wt/vol) trichloroacetic acidand stained for 30 min, after which the excess dye is removed by washingrepeatedly with 1% (vol/vol) acetic acid. The protein-bound dye isdissolved in 10 mM Tris base solution for OD determination at 510 nmusing a microplate reader. The results are typically linear over a20-fold range of cell numbers. CYD-2-11 has an IC₅₀ of 1.93 μM,Derivative CYD-2-17 has an IC₅₀ of 5.08 μM and CYD-2-13 has an IC₅₀ of5.91 μM. Data additional obtained from this assay is provided in FIG. 5.SMBA1 has an IC₅₀ of 7.35 and CYD-4-61,2-((3-((2-nitro-fluoren-9-ylidene)methyl)pyridin-2-yl)oxy)ethanamine hasan IC₅₀ of 0.026.

1. A compound of Formula I,

or salt thereof wherein,

is a double or single bond; A ring is a carbocyclyl, aryl, orheterocyclyl; X is CH or N; Y is (CH₂)_(n) or a direct bond to the Aring, wherein n is 1 or 2; R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are eachindividually and independently hydrogen, alkyl, halogen, nitro, cyano,hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, alkylthio,alkylamino, (alkyl)₂amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl,carbocyclyl, aryl, or heterocyclyl, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, andR⁸ are optionally substituted with one or more, the same or different,R¹⁰; R² is nitro or amino wherein R² is optionally substituted with oneor more, the same or different, R¹⁰; R⁹ is hydroxy, alkoxy, or amino,wherein R⁹ is optionally substituted with one or more, the same ordifferent, R¹⁰; R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino,mercapto, formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino,(alkyl)₂amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,aryl, or heterocyclyl, wherein R¹⁰ is optionally substituted with one ormore, the same or different, R¹¹; R¹¹ is alkyl, halogen, nitro, cyano,hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, alkylthio,alkylamino, (alkyl)₂amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl,carbocyclyl, aryl, or heterocyclyl, wherein R¹¹ is optionallysubstituted with one or more, the same or different, R¹²; R¹² ishalogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl,amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl,methoxy, ethoxy, acetyl, acetoxy, methylamino, ethylamino,dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino,N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio,methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,carbocyclyl, aryl, or heterocyclyl.
 2. The compound of claim 1, wherein

is a double bond.
 3. The compound of claim 1, wherein the A ring isphenyl ortho- or mata- or para-substituted with R⁹ wherein R⁹ ishydroxy, alkoxy, alkylamino, or substituted with hydroxy, (alkyl)₂amino,alkylsulfamoyl, dialkylsulfamoyl, or a heterocyclyl such aspyrrolidinyl, morpholinyl, piperazinyl, wherein heterocyclyl may besubstituted with one or more R¹².
 4. The compound of claim 1, wherein Yis a direct bond to the A ring.
 5. The compound claim 1, wherein X isCH.
 6. The compound claim 1, wherein the A ring is a heterocyclyl suchas pyridinyl ortho- or meta- or para-substituted with R⁹.
 7. Thecompound claim 1, wherein R² is nitro.
 8. The compound of claim 1,wherein Y is a direct bond to the A ring.
 9. The compound of claim 1,selected from the group:2-Methoxy-3-(2-nitro-fluoren-9-ylidenemethyl)-pyridine;9-(2-Methoxy-benzylidene)-2-nitro-9H-fluorene;2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol;2-Methoxy-3-(2-nitro-fluoren-9-ylidenemethyl)-pyridine;9-(2-Methoxy-benzylidene)-2-nitro-9H-fluorene;3-((2-Nitro-9H-fluoren-9-ylidene)methyl)pyridin-2(1H)-one;9-(2-Methoxy-benzylidene)-9H-fluoren-2-yl-amine;4-{2-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-morpholine;1-{2-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-piperazine;2-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethanol;2-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethylamine;4-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-piperidine;1-(4-Fluoro-benzenesulfonyl)-4-{2-[2-(2-nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-piperazine;1-(4-{2-[2-(2-Nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-piperazin-1-yl)-ethanone;1-Cyclopropanesulfonyl-4-{2-[2-(2-nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-piperazine;1-Methanesulfonyl-4-{2-[2-(2-nitro-fluoren-9-ylidenemethyl)-phenoxy]-ethyl}-piperazine;1-(4-Chloro-benzyl)-3-(2-nitro-fluoren-9-ylidenemethyl)-1H-indole; and2-((3-((2-nitro-fluoren-9-ylidene)methyl)pyridin-2-yl)oxy)ethanamine orsalts thereof.
 10. A pharmaceutical composition comprising a compound ofclaim 1, and a pharmaceutically acceptable excipient.
 11. Thepharmaceutical composition of claim 10, further comprising a secondtherapeutic agent.
 12. A method of treating or preventing cancercomprising administering a pharmaceutical composition of claim 10 to asubject diagnosed with, exhibiting symptoms of, or at risk of cancer.13. The method of claim 12, wherein the pharmaceutical compositions isadministered in combination with a second chemotherapeutic agent. 14.The method of claim 13, wherein the second chemotherapeutic agent isgefitinib, erlotinib, docetaxel, cis-platin, 5-fluorouracil,gemcitabine, tegafur, raltitrexed, methotrexate, cytosine arabinoside,hydroxyurea, adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin,idarubicin, mitomycin-C, dactinomycin and mithramycin, vincristine,vinblastine, vindesine, vinorelbine taxol, taxotere, etoposide,teniposide, amsacrine, topotecan, camptothecin bortezomib anegrilide,tamoxifen, toremifene, raloxifene, droloxifene, iodoxyfene fulvestrant,bicalutamide, flutamide, nilutamide, cyproterone, goserelin,leuprorelin, buserelin, megestrol anastrozole, letrozole, vorazole,exemestane, finasteride, marimastat, trastuzumab, cetuximab, dasatinib,imatinib, bevacizumab, combretastatin, thalidomide, and/or lenalidomideor combinations thereof.
 15. The method of claim 12, wherein the canceris selected from the group consisting of leukemia, cervical, ovarian,colon, breast, gastric, lung, skin, ovarian, pancreatic, prostate, head,neck, and renal cancer.
 16. Use of a compound as provided in claim 1, inthe production of a medicament for the treatment or prevention ofcancer.